Alexa Fluor® 594 anti-GFAP Antibody

Product Details

Verified Reactivity

Human, Mouse, Rat

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

Bovine spinal cord homogenate

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.

Preparation

The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 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

IHC-F - Quality tested
IHC-P - Verified
SB - Community verified

Recommended Usage

Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 1.0 - 5.0 µg/mL is suggested. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1.0 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.

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

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Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

Application References

1. McLendon RE and Bigner DD. 1994. Brain Pathol. 4:221.
2. Liu W, et al. 2011. Proteomics 11:3556. (FC) PubMed

Product Citations

1. Saito Y, et al. 2021. Front Immunol. 12:692321. PubMed
2. Radke J, et al. 2022. Nat Commun. 13:7304. PubMed
3. Watson A, et al. 2017. PLoS One. 10.1371/journal.pone.0180486. PubMed

RRID

AB_2566205 (BioLegend Cat. No. 644708)

Antigen Details

Structure

Around 50 kD

Distribution

Only expressed in astrocytes in the central nervous system.

Function

Together with other proteins to form intermediate filaments which supports astroglial cells. Astroglial cells support and nourish cells in the brain and spinal cord. If brain cells are injured through trauma or disease, astroglial cells react by rapidly prodcing more GFAP protein.

Cell Type

Astrocytes

Biology Area

Cell Biology, Neuroscience, Neuroscience Cell Markers

Molecular Family

Intermediate Filaments

Gene ID

2670 View all products for this Gene ID

UniProt

View information about GFAP on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Material Safety Data Sheet

Reviews

Related Protocols

• Immunohistochemistry Protocol for Frozen Sections
• Immunohistochemistry Protocol for Paraffin-Embedded Sections

Related Pages & Pathways

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.

Other Formats

Certificate of Analysis