Workable GAD 65/67 Antibody

Physiology
Jali Medical Inc.
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Rabbit anti-GAD 65/67

Catalog Number:

ab49832

I have tried many GAD 65, GAD 67 and combination antibodies to try to label inhibitory interneurons in perfused mouse brain tissue. This antibody is by no means the best but it does the job and provides sufficient staining for colocalization studies where I have used a second antibody.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse brain tissue interneurons

Primary Incubation

Slices were then quenched with 50 mM ammonium chloride for 15 minutes and washed with PBS

Blocking Agent

Blocked with a 5% Normal Goat Serum and 1% Bovine Serum Albumin in 0.1M PBS solution

Secondary Incubation

42.4 micrograms/mL for 24 hrs in 0.1 M PBS containing 1% NGS, 0.5% BSA, and 0.1% Tween-20 solution at 4 degrees

Tertiary Incubation

Donkey anti-rabbit Alexa 647 of 4µg/ml in 0.1M PBS for 1 h at room temperature

Detection

Fluorescence

Results Summary

The GAD 65/67 antibody is used to stain all interneurons in mouse brain slices. The stain is dependent on the quality of the perfusion as poor perfusion can lead to non-specific staining of certain structures. Overall, when combined with a second colocalizing stain this will provide some confirmation on whether the neuron stained is an interneuron. Shown image is at 60x using a confocal microscope.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Works better than most in the market

The Bad

Dependent on the perfusion

The Bottom Line

An OK antibody that is best used in combination with a second stain.

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