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Video - Simultaneous Proteomics and Genomics: TotalSeq and the Future of Single Cell Analysis
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Blog - ChIP Troubleshooting and Optimization
Chromatin immunoprecipitation (ChIP) is an important application to help determine where transcription factors bind or identify epigenetic changes at a gene of interest. As with most applications, understanding and using the right protocol is critical for success. But, when that protocol doesn’t work, it's hard to know where things have gone wrong. Here, we'll discuss 3 major areas to optimize or troubleshoot your ChIP experiments.




1. Chromatin Quality

The quality of the chromatin going into the IP is one of the most important aspects of a ChIP assay. Chromatin can degrade quickly, so ensure you always keep your samples on ice, and don't forget to add protease inhibitors during cell lysis. Once you've crosslinked and lysed your cells, there are two widely-used techniques for shearing chromatin: sonication and enzymatic digestion.


Overview of a ChIP experiment.
  • Enzymatic digestion utilizes an enzyme like micrococcal nuclease (MNase) which digests nucleic acids at a specific sequence. While this is a milder digestion compared to sonication, it does exhibit sequence bias.


  • Sonication uses mechanical shearing and can be advantageous when you are working with cells that are hard-to-lyse and unlike enzymatic digestion, it is an unbiased shearing method.


    • Probe sonicators use a probe placed directly in your sample. While this allows you to use shorter sonication times, it can also result in foaming of the sample and typically, you can only sonicate a single sample at a time. Because the probe is touching the sample directly, you need to ensure that you do not cross-contaminate your samples during the sonication.

    • Water bath or cup horn sonicators allow you to place all of your samples into a water bath that will indirectly deliver energy to your samples. These are ideal for sonicating multiple samples simultaneously.
So, which method is ideal for your experiments? This may depend on what protein you are pulling down or what cells you are using. If you're looking at an abundant, tightly bound protein like a histone, then either method should work well. If you are looking at a transcription factor, it may be better to use a gentler method like enzymatic digestion or try sonication using a lower concentration of detergent in the buffer. Cell types that are hard to lyse may require harsher sonication-based shearing protocols.


The exact conditions for shearing will depend on the specific cells and proteins you are analyzing. We'd recommend performing optimization experiments testing a variety of conditions before starting your IPs. Treat your cells exactly as you would for your experiment and run the resultant DNA on an agarose gel and look for shearing between 150-900 bp. With some cell types, even one extra cycle of sonication can cause over-shearing of your chromatin.



Chromatin from cell lines digested using MNase. On the left, chromatin was sheared optimally. On the right, chromatin was over-digested resulting in the majority of fragments below 400 bp. This can impair antibody binding resulting in little or no enrichment.

2. Immunoprecipitation
Once you’ve prepared the chromatin, the next step is to immunoprecipitate your protein of interest. Setting up the IP is a fairly simple process but there are a few things to keep in mind at this step.
  • Antibody Selection: Choose an antibody that has been validated for ChIP experiments (like our Go-ChIP-Grade™ antibodies). If this isn't possible, you can start by trying an antibody that has been validated for IP and be sure to use a positive control antibody and primers. Or, if there aren't any ChIP validated antibodies to your target, try expressing your protein with an epitope tag and using a ChIP-validated epitope tag antibody.


    • Some researchers prefer using polyclonal antibodies for ChIP because they can bind to multiple epitopes in case one or more epitopes were blocked during cross-linking. However, monoclonal antibodies will exhibit more lot-to-lot consistency and may be a better choice for long-term experiments.


  • IP Buffers: Some shearing protocols (like sonication) use a buffer containing high detergent concentrations; however, these same detergents can disrupt antibody binding. If you're using high detergents to shear, make sure that you prepare your IPs in a low detergent buffer to help dilute it out. Or, try a kit, like our Go-ChIP-Grade™ Protein G Enzymatic Kit, which contains all of the buffers you need for the experiment.


  • Quantity of Antibody and Chromatin: The exact amount of chromatin to add to any ChIP assay will vary based on the abundance of your protein of interest or the cell type being used. Typically, you can start with 3-5 μg of chromatin per IP, but you may need to scale up. Look to the manufacturer's recommendations to determine the optimal concentration of antibody to add.
3. Controls
Using the right controls allows you to properly analyze your downstream data at the end of the experiment. If something goes wrong, looking at how your controls performed will help you understand where to focus your troubleshooting efforts. Here, we'll outline the most important controls and how to use them.
  • Antibody Controls: Use a positive control antibody like one targeting RNA polymerase II or histones to ensure that you are using good quality chromatin and appropriate conditions. Include an isotype control to account for background in your assay and to measure relative enrichment of your protein at a target gene.


  • Input Controls: Save 1-5% of chromatin prior to the immunoprecipitation; this is known as the input control. It allows you calculate the amount of the enriched DNA relative to the total amount of DNA in the experiment. This is especially important if you are comparing enrichment between two samples that received different treatments.


Looking at ChIP data, you can see how each of the controls help analyze the data. Here we used an anti-HDAC1 antibody (A) or an isotype control antibody (B). For each, we measured enrichment at a positive control locus, CDKN1A, and a negative control gene locus, an α-Satellite region. Lastly, by using an input control, we presented the enrichment relative to signal from the input control, so that we can directly compare samples IP'ed with two different antibodies.
  • Primers: If you're performing qPCR analysis, design primers for a 100-250 bp region around the binding site of interest and include positive and negative control primers. A positive control primer is designed around a region that you know the protein binds. For this, look into the literature or see how the manufacturer validated the antibody you are using. Also include a negative control primer for a location where you wouldn't expect to see binding. These controls will help you gauge the levels of enrichment you'd expect to see at a true binding site for your protein.

Optimizing and troubleshooting your ChIP experiment can seem overwhelming, but breaking the experiment down can help understand where and why the experiment has failed. For more tips and tricks to help optimize your experiment, check out our ChIP webpage or watch our ChIP protocol video.

Image by Dzu-Doodles.
Contributed by Kelsey Swartz, Ph.D.


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Blog - BioLegend at IMMUNOLOGY 2019™
Immunologists will soon be gathering for one of the largest immunology-focused conferences of the year. The American Association of Immunologists holds an annual meeting, with this year’s taking place May 9-13 in our hometown, San Diego. The conference is a chance for researchers to gather, collaborate, and learn about the latest scientific findings. Given its importance, BioLegend has served as its top platinum sponsor since 2008. In addition, this year marks an important milestone as we move into our new 8-acre campus, equipped with state-of-the-art technology and amenities, allowing us to employ over 1,000 employees in a single location.

Aside from that, let’s go over some of the biggest events, products, and promotions that we’ll be featuring at IMMUNOLOGY 2019™!





Meet the Experts!
We’ll be at booth #901, where you can talk to any member of our team. If you’d like to get deeper into the science, seek out one of our exhibitor workshops where we’ll discuss spectral flow cytometry and the benefits of TotalSeq™, our oligonucleotide-conjugated antibodies for proteogenomics studies. Each workshop will give away three $50 Amazon gift cards, so be sure to attend! We’ll also have a number of presenters at poster sessions, covering topics ranging from signaling pathways and novel antibody clones to new LEGENDplex™ kits. And, if LEGENDplex™ sparks your interest, be sure to get a hands-on demo with our new cloud-based software to help with your multiplexing analysis.


BioLegend Supports the Research Community
As a supporter of immunologists everywhere, we also sponsor several AAI Awards, including the AAI-BioLegend Herzenberg Award, named in honor of Leonard Herzenberg who made innumerable contributions to the fields of flow cytometry and immunology. Winners of this award have made outstanding contributions to B cell research. This year’s award recipient is Frederick W. Alt, Ph.D. of HHMI, Boston Children’s Hospital. Alt’s lab focuses on B cell antigen receptor repertoires. We also sponsor the AAI-Lefrançois-BioLegend Award, in honor of Leo Lefrançois, to provide a travel award to an outstanding graduate student or post-doc in the mucosal immunology research field. This year’s winner is Scott M. Anthony, Ph.D. of the University of Iowa. His work focuses on resident memory CD8+ T cells.

Frederick W. Alt, winner of this year’s AAI-BioLegend Herzenberg Award. Image from HHMI.
 
Purple Perks
It wouldn’t be a conference without giveaways and promotional items, so make sure you stop by our booth! You’ll be able to spin the wheel for prizes like our Rock the Synapse T-shirt, a Plexy Plush doll, Starbucks gift cards, and more! And, you can learn more about our additional promotions, like our Make the Switch campaign, where you can win a Nintendo Switch™ with the purchase of recombinant proteins, like cytokines or growth factors. You can also pick up some groovy stickers to help decorate your MojoSort™ magnet for cell separation.






If you visit the BioLegend booth, you’re sure to leave with lots of swag.
Festivities
In addition to the Service Appreciation Reception (by invitation only), we’ll be sponsoring the IMMUNOLOGY 2019™ Gala, which will take place on the USS Midway Museum. The Midway is an iconic naval craft carrier that was in service from 1945 to 1992. It now serves as a museum that houses a variety of restored naval aircraft, helicopters, and more. We look forward to seeing you out there!

The USS Midway is an impressive sight and will host the IMMUNOLOGY 2019™ Gala. Image from Visit California.
San Diego Fun Facts
As you visit our hometown, we hope you enjoy the conference and sights that San Diego has to offer. Here are some fun facts to get you excited for your trip:
  • The film “Some Like It Hot”, starring Marilyn Monroe, was filmed at the Hotel del Coronado in 1958.
  • Balboa Park is actually larger than New York’s Central Park and houses 17 different museums, many of which offer free admission.
  • San Diego produces more avocados than any other city in the country.
  • In 1969, Ronald Reagan was the first person to drive across the newly opened Coronado Bridge.
  • San Diego has inspired a number of songs, including: Bing Crosby’s “Where the Turf Meets the Surf,” Bruce Springsteen’s “Balboa Park,” and Eric Clapton’s “The Road to Escondido.”
 
Contributed by Ken Lau, Ph.D.


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Blog - Keeping it Simple with Fluorescence

Several applications utilize immunofluorescence and fluorescently conjugated antibodies. This blog focuses specifically on fluorescence microscopy, including immunohistochemistry and immunocytochemistry assays. Fluorescent detection allows you to look at multiple markers in a single sample by using discrete excitation sources for each fluorophore. This can be performed directly with a fluorophore-conjugated primary antibody or indirectly with an unconjugated primary and a fluorophore-conjugated secondary antibody. Here, we'll discuss the benefits of direct conjugates and how they can help simplify your microscopy experiments and allow you to easily expand your multicolor panels.

Microscopy is commonly used when you want to visualize proteins in their native environment. As more advanced microscopy techniques are always being developed, there are many ways you can use microscopy in your research. We've listed some of the most common reasons below.


1.) Protein-to-protein interactions can be analyzed in their native environment.

This is often beneficial, as most assays tend to "mimic" the native environment of the cells of interest. This can lead to faulty conclusions and inconclusive results.

2.) Protein localization can also be determined through microscopy.

This too, can play an important role, as proteins function differently in distinct parts of the cell. In addition, proteins can be quantified in those specific environments. Again, this plays an important role, especially when a treatment is applied to an organism, and understanding where that protein is increased.


The Simpsons, 20th Century Fox Television




3.) Lastly, these microscopy samples can be saved for longer periods. This provides flexibility to longitudinal studies and clinical trials where researchers can re-stain samples. Given these reasons, fluorescence microscopy is both broad and flexible.
While there are many different forms of microscopy, fluorescence microscopy bifurcates into two categories, which are separated by their respective sample types.

Samples that are derived from cells are considered immunocytochemistry (cyto=cell). Typically, these cells are either grown on or spun onto a slide and stained with either just a primary antibody or a primary and secondary antibody for signal amplification.

The other major category involves a whole tissue section. These whole tissues can be sectioned at varying depths (depending on design and intention of the assay). These samples are then referred to as immunohistochemistry, as they utilize tissue sections (histo=tissue section) to study the sample of interest.



Mouse brain stained with Alexa Fluor® 594 anti-Tubulin β3, Alexa Fluor® 647 anti-MAP2, and DAPI.


Traditionally, these methodologies rely on the use of a primary and a secondary antibody. The secondary antibody will either have a fluorophore conjugated to the antibody or will use an alternative read-out method such as HRP. While the BioLegend catalog currently offers a great deal of purified antibodies for both neuroscience and cell biology applications that involve microscopy, we also have great directly conjugated primary antibodies against a variety of targets.


While it is true that signal amplification is greatly increased when using a two-step stain, there are some advantages of using a directly conjugated primary antibody that go overlooked.

1.) Non-specific binding will always increase when you add more antibodies into an application.

Antibodies display some degree of non-specific binding, regardless of specificity. If you only require one primary antibody, you might find that you produce a cleaner image.

2.) The experiment itself will be simpler and more streamlined.

We all know long days in the lab do not necessarily generate effective and consistent results. Thus, with directly primary antibodies, you can help reduce experimental error and save yourself time.

3.) The use of single-step stain also makes it significantly easier to multiplex at a higher rate without worrying about whether your secondary reagent will cross-react to multiple primary antibodies that share the same isotype. This can help make your experiment more effective.

4.) Whenever our team develops a new clone and/or is interested in conjugating a new fluorophore to a primary antibody, we perform a side-by-side comparison of both the single-step stain and the two-step stain to ensure the directly conjugated primary antibody stains the target equally or brighter than the purified antibody before that antibody is placed in our catalog.





Mouse brain stained with Alexa Fluor® 488 anti-Myelin Basic Protein, Biotin anti-IP3R1, Alexa Fluor® 594 anti-MECP2, and DAPI. HeLa cells stained with Alexa Fluor® 488 Vimentin, Alexa Fluor® 594 Cytochrome C, and DAPI.


Ultimately, the decision to use a two-step stain versus a one-step stain will always be assay-dependent as my colleague pointed out in her blog discussing primary vs secondary detection systems.

For these reasons, we continue to grow our product lines for both neuroscience and cell biology targets for primary antibodies that are both affordable and effective. To learn more about direct conjugates in microscopy, check out our bio-bit.


Contributed by Sean Cosgriff.


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Blog - Primary vs Secondary Detection Systems

What is a Secondary?
A secondary reagent is used to identify or enable the detection of a primary antibody or reagent. Primary reagents are antibodies specific to the target or cell marker of interest. If the primary antibody is unlabeled, a secondary reagent that is labeled would be required to bind to the primary reagent on the cell and enable its detection.

While many secondary detection systems use a purified antibody with an anti-Ig secondary antibody, biotin-labeled antibodies and conjugated streptavidin molecules can also be used. Biotin is a water-soluble form of vitamin B that is used as a cofactor for a variety of metabolic processes. Streptavidin is a bacteria-derived protein with an extremely high affinity for biotin (Kd of approximately 10-14). These two molecules bind together almost irreversibly, making them ideal candidates for secondary detection.


Staining is not a Highlander situation. You can use more than one staining reagent, and sometimes it is actually better!

Highlander, Cannon Films.


Advantages of Using Secondary Reagents
There are 3 main reasons why you would want to choose a secondary labeling system for your experiment.

1. Restrictions on Formats of Primary Antibody Available

Sometimes for studies, researchers will need to have an antibody in a specific fluor or format that may not be commercially available. It can be easier to find a purified or biotinylated version of the primary antibody of choice. In contrast, secondary reagents are very flexible and are often available in a wider variety of formats than primary antibodies.

2. Expression Levels of Target are Low

Using a secondary reagent for detection often leads to signal amplification that can make low-expressing targets easier to detect. Many secondary reagents are conjugated at similar fluorophore to protein ratios as primary antibodies, but it is common for multiple secondary antibodies to bind to a single primary antibody, giving an overall stronger signal.



Comparing the signal detection of a primary labeling system to signal from a primary + secondary labeling system

3. Reagent Flexibility

Incorporating a secondary reagent can allow a single primary antibody to be used in multiple applications. For example, if you have an antibody that works for western blotting and microscopy, utilizing secondary reagents can allow you to use this clone for both applications. Secondary reagents are often more economical than buying additional directly conjugated antibodies and can add flexibility in panel selection for flow cytometry. For example, purchasing biotinylated lineage antibodies can allow you to easily adjust the color of your lineage cocktail by simply swapping out the streptavidin used to label the lineage markers rather than replacing the entire cocktail.


How to Choose the Correct Secondary Reagent
There’s a few factors that need to be taken into consideration when choosing the best secondary reagent for your experiment.


1. Conjugation

It's important to verify that the secondary antibody has the proper conjugation for your experiment. If you are doing a western blot, an HRP conjugate can be used with a substrate solution, whereas a fluorophore-conjugated secondary is more appropriate for use in flow cytometry. While the majority of our fluorophores can be used for flow, we recommend using the Alexa Fluor® dyes, Brilliant Violet™ 421, and Brilliant Violet™ 510 for microscopy applications. Check out our microscopy page to see how these products perform in ICC, IHC-F, and IHC-P.

2. Primary Antibody Host Species & Sample Type

The most commonly used secondary reagents are antibodies that are raised against the immunoglobulins of another species, so you will need to check the host species and isotype of your primary antibody to ensure your secondary will recognize the primary antibody. You'll also need to keep your sample type in mind; for example, using an anti-mouse secondary on mouse samples may result in high background as the secondary will recognize both the primary antibody of interest as well as any endogenous immunoglobulins that may be present in the sample.

3. Staining Panel Composition

When staining with multiple primary antibodies, it is important to ensure that your secondary reagent will only bind with one primary antibody of interest. This can sometimes be difficult to do as antibody availability can be limited, so consider using a biotin-labeled primary with a streptavidin-labeled secondary to help mitigate this issue. For larger multicolor panels, it may be necessary to use directly conjugated primary antibodies instead of secondary antibodies.

4. Assay Adjustments

Utilizing secondary reagents can also impact how you design your overall experiment. Incorporating secondary reagents can sometimes lead to increased non-specific binding and background noise. As such, it is important to include a secondary-only stained sample as a control. Additional blocking steps may also be needed. For secondary antibodies, blocking with a buffer containing serum that matches the species of the secondary can help reduce non-specific binding to the surface of your cells. Using streptavidin secondaries may require a protocol to block or neutralize any endogenous biotin that may be present in your sample.


Still not sure if you should use a direct conjugate or secondary system? Check out our secondary reagents or email us at tech@biolegend.com and we'll help you choose!


Contributed by Samantha Stanley, PhD.


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Podcast Episode - Anti-Vaxxers and the Quest for Measles
In this latest podcast, we tackle the resurgence of Measles, vaccinations, and common anti-vaccination myths!

Note regarding our talk on the polio vaccine: Following its mass production in 1955, certain pharmaceutical companies failed to properly inactivate the Salk vaccine, leading to over 250 reported cases of paralysis. Salk’s inactivated polio vaccine was the first developed, followed by the oral polio vaccine (OPV). OPV remained in use in the US until 1999 where it was replaced with an inactivated, injectable vaccine. However, OPV is still used in other countries around the world.

Topics

Common vaccination myths
WHO's ten threats to global health in 2019
Teen secretly gets vaccinated, speaks to Senate
Unvaccinated teen sues school for limiting his activity
Facebook to ban anti-vaxx ads in new push against 'vaccine hoaxes'
Measles returns to Costa Rica thanks to unvaccinated family
Anti-Vaxx mom asks how she can protect her 3 year old from Measles outbreak
Study on 657,461 children finds no link between autism and vaccines

Keywords: vaccines, measles, immunology, anti-vaxxers, autism, Andrew Wakefield, myths, science, Ethan Lindenberger, kale, global health, chickenpox


Having difficulty playing this episode? Listen on PodBean


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Blog - It’s Not All About the Antibodies


The irreproducibility of published data has become a legitimate concern in recent times. In 2012, a landmark study conducted by Glenn Begley and his team concluded that the majority of cancer research studies were not reproducible, demonstrating that a startling 47 out of 53 published findings could not be replicated (Nature 483, 531-533; 2012). The implications for biomedical research are potentially staggering considering the billions spent on research every year, with some suggesting that as much as $28 billion is wasted on irreproducible pre-clinical research every year (Freedman LP, et. al. PLoS Biology 13: e1002165). Antibodies used in research have become a popular target for blame. Many instances of poor specificity, cross-reactivity, and lot-to-lot variability have cost researchers millions and possibly billions, and will likely continue to do so in the future. But, we need to have a keen eye for other factors that are likely to contribute to experimental irreproducibility beyond just the antibodies.
Bias
As humans, we all have preferences, desires, pressures, and feelings that might be reflected in the experimental outcomes, no matter how hard we try to separate them from data. One of the difficulties with a hypothesis-based approach is that most researchers want the data to support their original hypothesis. This inherently introduces bias from the start. Along the way, bias contributes to almost all aspects of experimentation. Below is a list of some areas, but not all, where bias has effects:
  • Selective reporting, i.e. excluding outlier data
  • Data selecting, i.e. counting phenotypes by microscopy
  • Limited data sets
  • Interpretation of data
  • Experimental design
  • Reagent selection
  • Collaborators
There are a few straightforward steps that one might take to try to remove bias as much as possible. These might include:
  • Blinding and randomization of treated versus control groups
  • Including as many controls as possible, both positive and negative
  • Use proper statistics to determine significance
  • Use well published protocols and reagents
  • Use quantifiable measurements, making sure all instruments and assays are properly calibrated


Comic from Dzu-Doodles.

Variability
Commercial resources for critical research reagents like cell culture media, antibodies, and recombinant proteins may go through many rounds of validation testing prior to sale, but what about other day to day materials that are commonly used? Some things to consider:
  • Do you routinely check the quality of your DI water?
  • Are any of your chemicals expired?
  • When was the last time your instruments and equipment were calibrated?
  • What is the range of temperature fluctuation in your refrigerator?
  • Do you always buy the same reagents from the same source? Even if you do, how does the supplier control lot-to-lot variability?
  • How long do you keep stock solutions? Would you know if they are contaminated?
  • Are conditions for the storage and handling of mice consistent from week to week, and month-to-month?
  • Are your cell lines authenticated? Are they mycoplasma free?


Comic from Dzu-Doodles.

The potential for variability and irreproducibility, even within the same lab, are likely too high if processes, reagents, equipment, and protocols are not consistent.


In addition to variability in research materials, there is also an inherent variability in human and research animal subjects that makes reproducibility a challenge.


Trainees
The academic system relies on a large proportion of its data to be generated by people regarded as "trainees," either graduate students or post-docs, while experienced Principal Investigators spend their time seeking funding and writing grants. Their status as trainees reflects on their lack of expertise in a subject matter or experimentation, often times both. Yet, the expectation is that they perform the majority of the experiments to generate data for publications. Even in labs with the most well renowned PI's, there will always be new trainees entering, and creating new data. In addition to this, experimental training comes in vastly different forms and quality. Some students are trained by experienced mentors, while others may be trained by the student who started in the lab last quarter. The lack of universal standards for training should be major concern for reproducibility in science.


Comic from Dzu-Doodles.



Conclusion
It's not all doom and gloom though. Scientific discoveries have led to multitudes of breakthroughs that have directly impacted the quality and quantity of life we get to have today. New discoveries will be built upon the groundwork of science that came before. Despite all the imperfections in the way we do science, the body of work that science builds upon provides us a means of constantly changing our understanding of the world and ourselves. We do have a long way to go in reducing irreproducibility, but just consider: it's not always the antibodies.


Comic from Dzu-Doodles.

Learn how BioLegend addresses Validation and Reproducibility.
Written by Dzung Nguyen, PhD.


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Video - Isolate PBMCs with Lymphopure™
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Blog - Increase Your Knowledge: Learn how BioLegend helps you stretch your neuro-dollar further.



Know what you’re working with
Reproducibility is the cornerstone of science. Using a consistent amount of antibody from experiment to experiment is crucial to ensure the reproducibility of your data. BioLegend offers three sizing options: microgram, microliter, and test sizes. Understanding the differences between the formats can help you decide which format will best suit your needs. Microgram sizing is perhaps the best way to know exactly how much antibody you are buying, and is often the best value. Similarly, microliter sizes are sold at a set volume with the concentration clearly stated on the TDS. Both the microgram and microliter sizing options are best suited for optimizing titration, and allow for consistent antibody usage across experiments. They also make it easier to calculate how much isotype control to use. Test sizes are offered as a convenience, where the titering has already been done for you. Test size antibodies are meant to be used at a set volume (typically 5 μL/test).




Discover more, for less
Technical and biological replicates ensure sound and reproducible results, but come at the cost of increased reagent usage. BioLegend offers competitive pricing on our neuroscience products, so you get more replicates and more data. Check out the table below to see how you can get more value in your vial when buying the same antibody clone from BioLegend and a few of our competitors. For ease of comparison, we converted the size of those products sold in a 100 μL format (marked with an asterisk) to total μg per vial based on their concentration of 1 mg/mL.



Description Clone BioLegend
(total μg/vial)
Competitor A Competitor B
α-Synuclein Phospho
(Ser129)
P-Syn/81A 100 μg*- $285 50 μl- $419 100 μg- $333
Brevican N294A/6 100 µg*- $140   100 µg- $353
GluR2 L21/32 100 μg*- $140 100 μg- $419 100 μg- $353
GRK N145/20 100 μg*- $140 100 μg- $405 100 μg- $353
KCC2 N1/12 100 μg*- $140 100 μg- $409 100 μg- $353
Laforin N84/37 100 μg- $140 100 μg- $389 100 μg- $343
MECP2 N227/21 100 μg*- $180   100 μg- $343
NeuN 1B7 100 μg*- $202 100 μg- $429  
Pan-shank N23B/49 100 μg- $140 100 μg- $409 100 μg- $353
VDAC1 N152B/23 100 μg*- $140 100 μg- $405 100 μg- $343


One Product, Multiple Applications
Here at BioLegend, we quality test and validate our products across a wide range of applications. Many of our reagents pull double, triple, or quadruple duty, meaning you can perform more assays with the same reagent. These applications are listed on the TDS so you know what your shiny new antibody can do.
Purified anti-Tubulin β 3 (TUBB3) Clone TUJ1




Particular application not listed? Even if we don’t test a clone for a particular assay in-house, our dedicated team of technical support scientists scour the literature for other possible applications for our clones so you don’t have to.



Generous Pricing and Discounts
You can learn more about our neuroscience products with our helpful webpage. For discounts, quotes, and help with our products, get in touch with our technical application scientists.

Contributed by Christopher Dougher, PhD.


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