Iba1 in Rat Brain

December 21, 2015

BlueskyReddit
Pharmacology
Brody School of Medicine
PhD student

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Company:

Wako

Product Name:

Wako anti Iba-1, rabbit polyclonal

Catalog Number:

019-19741

Anti Iba1 antibody from Wako was used to label microglia in rat brain areas associated with reward and memory such as the hippocampus, nucleus accumbens, and prefrontal cortex. This antibody was chosen as the antibody for staining microglia from Santa Cruz (FL147) failed.

Experimental Design and Results Summary

Applications

Immunohistochemistry

Sample

Rat brain sections

Primary Incubation

1:10,000 primary dilution in PBS containg 0.3%tritonx, 0.1%BSA, 1%NGS, 48-hours at 4OC on rocking platform. Free floating sections.

Blocking Agent

5% NGS in PBS containg 0.3% Triton x for 1 hour at room temperature

Secondary Incubation

Biotinylated ABC secondary PK4000 Vector

Tertiary Incubation

NA

Detection

DAB with 0.015% H2O2

Results Summary

Get robust staining all over the brain. Easy to detect microglia on 40um free floating sections. 48 hours was necessary to get a strong signal if using the PK4000 Vector kit.

Additional Notes

Remember to use goat anti rabbit secondary reagens. I recommend the one from Vector labs PK4000.

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Summary

The Good

Most widely accepted antibody for staining microglia. Get good staining with the ABC kits. Makes it easy to quantify microglia activation by morphology.

The Bad

Have to keep in freezer, so aliquot the antibody. Can react with monocytes. Also can't tell the polarization of the microglia.

The Bottom Line

Best one for immunostaining.

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