Pacific Blue™ anti-mouse CD21/CD35 (CR2/CR1) Antibody

Pricing & Availability
Clone
7E9 (See other available formats)
Regulatory Status
RUO
Other Names
CR2/CR1
Isotype
Rat IgG2a, κ
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Product Citations
publications
1_7E9_Blue_081908.jpg
C57BL/6 splenocytes stained with 7E9 Pacific Blue™
  • 1_7E9_Blue_081908.jpg
    C57BL/6 splenocytes stained with 7E9 Pacific Blue™
  • 2_66_Mouse_Lymph_Node_CD21_Pax5
    Mice were injected subcutaneously with sheep red blood cells in a volume of 25 µl per site on days 0 and 4 and harvested on day 11. Confocal image of C57BL/6 mouse lymph node acquired using the IBEX method of highly multiplexed antibody-based imaging: CD21 (purple) in Cycle 7 and Pax5 (cyan) in Cycle 8. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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123413 25 µg 104€
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123414 100 µg 235€
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Description

CD21, also known as CR2 (complement receptor 2) and C3d receptor, binds C3d and iC3b. It is also a receptor of Epstein-Barr virus. CD35, also known as CR1, binds C3b, iC3b, C4b, and iC4b. CD21/CD35 is primarily expressed on B lymphocytes, mast cells, follicular dendritic cells, macrophages, and activated granulocytes. CD21/CD35 forms part of the B-cell antigen receptor complex with CD19 and CD81 and is involved in signal transduction.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
CD35/CFA
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested

SB - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 1.0 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Violet Laser (405 nm)
Application Notes

Additional reported application (for relevant formats) include: spatial biology (IBEX)5,6.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

(PubMed link indicates BioLegend citation)
  1. Boackle S, et al. 2001 Immunity 15:775.
  2. de Andres B, et al. 2012. J. Immunol. 189:2300. PubMed
  3. Chiu YK, et al. 2014. J Immunol. 193:2207. PubMed
  4. Koening PA, et al. 2014. J Biol Chem. 289:34490. PubMed
  5. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Bhattacharya Y 2014. J Exp Med. 211:841. PubMed
  2. Farsakoglu Y et al. 2019. Cell reports. 26(9):2307-2315 . PubMed
  3. Menzel L, et al. 2021. Cell Rep. 37:109878. PubMed
  4. Juul-Madsen K, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  5. Dubey LK, et al. 2019. Cell Rep. 27:2442. PubMed
  6. Avery DT, et al. 2018. J Exp Med. 215:2073. PubMed
  7. Duan L, et al. 2021. Immunity. 54:2256. PubMed
  8. Onodera T, et al. 2016. J Immunol. 196: 4172 - 4184. PubMed
  9. Duong B, et al. 2010. J Exp Med. 207:173. PubMed
  10. Noviski M, et al. 2018. Elife. 7:e35074. PubMed
  11. Lofano G, et al. 2015. J Immunol. 195: 1617-1627. PubMed
  12. Simoni L, et al. 2020. Cell Rep. 33:108330. PubMed
  13. Seeley-Fallen M, et al. 2014. Proc Natl Acad Sci U S A. 111:9881. PubMed
  14. Radice E, et al. 2020. Cell Reports. 32(5):107951. PubMed
  15. Gill R, et al. 2017. Toxicol Appl Pharmacol. 330:22. PubMed
RRID
AB_2085159 (BioLegend Cat. No. 123413)
AB_2085158 (BioLegend Cat. No. 123414)

Antigen Details

Structure
145 kD/190 kD type I transmembrane glycoprotein
Distribution

B cells, follicular dendritic cells

Function
Signal transduction
Ligand/Receptor
Associated with CD19/CD81. CD21 binds C3d, iC3b, and EBV; CD35 binds C3b, iC3b, C4b, and iC4b.
Cell Type
B cells, Dendritic cells
Biology Area
Cell Biology, Costimulatory Molecules, Immunology, Neuroinflammation, Neuroscience
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References

1. Kozono Y, et al. 1998. J. Immunol. 160:1562.
2. Shimizu I, et al. 2007. Blood 109:1773.
3. Roozendaal R and MC. Carroll. 2007. Immunol. Rev. 219:157.

Gene ID
12902 View all products for this Gene ID 12946 View all products for this Gene ID
UniProt
View information about CD21/35 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

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For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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