Anti-GFAP as an Astrocyte Marker (for IHC/IF) from EMD Millipore

Behavioral Genetics Lab
Beckman Institute for Advanced Science & Technology
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Company:

EMD Millipore

Product Name:

Anti-Glial Fibrillary Acidic Protein, clone GA5

Catalog Number:

MAB360

This antibody was used in a study that investigated the crossover of astrocytes with aromatase in the teleost brain. It has not yet been published as of this date. Several anti-GFAP antibodies were tested in this study for various applications; the antibody reviewed here (MAB360) gave best results in IF staining and one of the best in IHC staining.

Experimental Design and Results Summary

Applications

Immunohistochemistry

Sample

Teleost brain tissue, 4% PFA postfixed, slide mounted

Primary Incubation

Anti-GFAP (Millipore, MAB360) incubated overnight at 4°C in blocking reagent (1:500 dilution)

Blocking Agent

5% Normal Goat Serum, 0.3% Triton-X in PBS

Secondary Incubation

Goat anti-mouse IgG (Jackson Immuno Research Labs, 115-005-003), incubated 2 h at RT (1:500)

Tertiary Incubation

NA

Detection

Avidin/Biotin Complex for 1h RT followed by DAB

Results Summary

The antibody worked excellently in chromogenic visualization of astrocyte cells, with no background staining compared to isotype control and adequate staining of processes in comparison to cell bodies.

Additional Notes

None

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Summary

The Good

High specificity and relatively high dilution

The Bad

Requires overnight incubation instead of 1h RT in order to visualize processes, could not validate that the antibody did not stain ependymal cells

The Bottom Line

This is a very good antibody to detect GFAP/astrocytes, and it seem to work very well in the telost brain, of which many antibody are not readily available. That being said, this antibody may not work in other teleost species (we use A. ocellaris), and the protocol may not cross over to other species. Remember to always optimize to your specific needs.

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