Santa Cruz Biotechnology, Inc.
Neu (3B5)
sc-33684
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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.
Immunofluorescence
Cytospins of human breast cancer cell lines: BT474 (HER2-high) and MDA-MB231 (HER2-low)
2ug/ml of Neu (3B5): sc-33684 in PBS-Tween buffer at room temperature for one hour.
None
Vector Dylight 549 anti-mouse antibody (D12549) used at 5.0 ug/ml, Room Temperature for 45 minutes
Vectashield mounting medium containing DAPI counterstain
Fluorescence microscopy- Nikon microscope with SPOT camera
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.
Very easy to optimize; publication-quality results
None noted
Overall, this mouse monoclonal for HER2/Neu is very useful for immunocytochemistry. We will definitely use it for future experiments.