Immunocytochemistry of a Her2 Variant

July 30, 2015

Center for Hearing & Deafness
University at Buffalo
Principal Investigator

Overall

Quality of Results

Ease-of-Optimization

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Company:

R&D Systems/Biotechne

Product Name:

ErbB2/Her2 (Herstatin variant)

Catalog Number:

MAB11291

Info: View Product View Product Specs

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We are studying Her2 expression in a number of cancer cell lines that may express Her2 variants. We selected this antibody because it reacts with a Her2 variant called Herstatin. We tested this antibody on cytospins of BT474 human breast cancer cells for optimization.

Experimental Design and Results Summary

Applications

Immunohistochemistry

Sample

BT474 human breast cancer cells prepared as cytospins on slides

Primary Incubation

1:20 dilution of MAB11291 at room temperature for one hour.

Blocking Agent

Used Scytek IHC kit: H2O2 block ADA008 (5 min) and Superblock AAA008 (4 min) room temp.

Secondary Incubation

Scytek IHC kit: ABZ008 Ready-to-Use

Tertiary Incubation

NA

Detection

DAB from Scytek Kit: 2X5 mins incubation.

Results Summary

We saw very intense DAB staining in the BT474 sample and a very clean negative control slide.

Additional Notes

PBS washes 3X between each step. Negative control received an isotope control antibody.

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Summary

The Good

This antibody worked very well at the concentration recommended by the manufacturer.

The Bad

The 1:20 dilution means that there is not a lot of room for increasing the concentration, if necessary.

The Bottom Line

Overall, this antibody worked very well for immunocytochemistry on cytospins of BT474 cells at the 1:20 dilution.

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