Good GAD65 Antibody for Immunohistochemistry

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Company:

Abcam

Product Name:

Mouse anti-GAD65

Catalog Number:

ab26113

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.  To this end, I wanted to identify GAD65 location in GABAergic cells in fixed brain tissue. This antibody worked well at a low dilution and identified the processes of the GAD65-positive cells, but not the cell bodies.

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

The cell processes stained, but the cell bodies did not. The dilution that gave the best pictures was much lower than the dilution recommended by the manufacturer. Worked better for IHC than for immunoblotting, but gave some response for both techniques.

Additional Notes

None

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Summary

The Good

Antibody works and gives good pictures.

The Bad

Can be more expensive than other companies' equivalents.

The Bottom Line

Worked well for IHC, would not as readily recommend for immunoblot.

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