HER2/Neu MAb From Santa Cruz For Immunocytochemistry

Center for Hearing & Deafness
University at Buffalo
Principal Investigator

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Santa Cruz Biotechnology, Inc.

Product Name:

Neu (3B5)

Catalog Number:

sc-33684

We are studying HER2 expression in retinoblastoma and need a variety of HER2 antibodies for our immunocytochemistry studies.  This particular HER2/Neu antibody from Santa Cruz is a mouse monoclonal. Most of our other HER2 antibodies are rabbit polyclonal antibodies, so this mouse monoclonal was chosen as an option for double-labeling studies to pair with other rabbit polyclonal antibodies.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Cytospins of human breast cancer cell lines: BT474 (HER2-high) and MDA-MB231 (HER2-low)

Primary Incubation

2ug/ml of Neu (3B5): sc-33684 in PBS-Tween buffer at room temperature for one hour.

Blocking Agent

None

Secondary Incubation

Vector Dylight 549 anti-mouse antibody (D12549) used at 5.0 ug/ml, Room Temperature for 45 minutes

Tertiary Incubation

Vectashield mounting medium containing DAPI counterstain

Detection

Fluorescence microscopy- Nikon microscope with SPOT camera

Results Summary

The results show very specific and intense staining for HER2 (red) in the BT474 breast cancer cell line (HER2-high). Very little immunoreactivity is seen in the MDA-MB231 cell line (HER2-low). The negative control shows only the DAPI counterstain (blue) for nuclei. We were very pleased with these results.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Very easy to optimize; publication-quality results

The Bad

None noted

The Bottom Line

Overall, this mouse monoclonal for HER2/Neu is very useful for immunocytochemistry. We will definitely use it for future experiments.

Share your experience with other scientists. Write a Review! »

Join the discussion