GAD67 Antibody Works Well for Immunohistochemistry

Biological Science
Florida State University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Chemicon (now Millipore)

Product Name:

Mouse anti-GAD67

Catalog Number:

MAB5406

In these experiments, I wanted to investigate whether a novel receptor (for olfactory bulb tissue) was co-expressed with known subpopulations. The antibodies used target known histological markers for these subpopulations.  Here, I wanted to determine GAD67 location in fixed brain tissue. This antibody had been left in the refrigerator for years before I used it. It worked well but labeled only the processes, not the cell bodies, of GAD67-positive cells.

Experimental Design and Results Summary

Applications

Immunohistochemistry

Sample

PFA-fixed brain tissue slice

Primary Incubation

1:10,000 for 48 hours at 4 degrees C

Blocking Agent

1% normal horse serum in PBS (0.4% Triton-X 100)

Secondary Incubation

1:1000 Cy3-conjugated horse anti-mouse for 2 hours at room temperature

Tertiary Incubation

NA

Detection

Cy3 fluorescence via fluorescent confocal microscopy

Results Summary

Cell processes were bright and easily identified. Staining occurred throughout the slice, as GAD67 is known to be more prominent than GAD65. Staining did not occur in lamina known to not express GAD65 or GAD67.

Additional Notes

Can be used at a much more dilute concentration than the company states.

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Summary

The Good

Works for a long time at a good dilution.

The Bad

Takes a bit of fine-tuning to get the dilutions correct.

The Bottom Line

A good antibody for IHC but not for immunoblot. Recommended.

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