Immunohistochemistry analysis of HER-2 / ErbB2 showing staining in the membrane of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a HER-2 / ErbB2 Rabbit Polyclonal Antibody (PA516774) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from C-terminus of human HER-2 protein. This sequence is identical in rat neu protein.|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunoprecipitation (IP)||See 1 publications below|
PA5-16774 targets HER-2 in IP and WB applications and shows reactivity with Human, mouse, Non-human primate, and Rat samples.
The PA5-16774 immunogen is a synthetic peptide from C-terminus of human HER-2 protein. This sequence is identical in rat neu protein.
This gene encodes a member of the epidermal growth factor (EGF) receptor family of receptor tyrosine kinases. This protein has no ligand binding domain of its own and therefore cannot bind growth factors. However, it does bind tightly to other ligand-bound EGF receptor family members to form a heterodimer, stabilizing ligand binding and enhancing kinase-mediated activation of downstream signalling pathways, such as those involving mitogen-activated protein kinase and phosphatidylinositol-3 kinase. Allelic variations at amino acid positions 654 and 655 of isoform a (positions 624 and 625 of isoform b) have been reported, with the most common allele, Ile654/Ile655, shown here. Amplification and/or overexpression of this gene has been reported in numerous cancers, including breast and ovarian tumors. Alternative splicing results in several additional transcript variants, some encoding different isoforms and others that have not been fully characterized.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
erbB3 is an active tyrosine kinase capable of homo- and heterointeractions.
PA5-16774 was used in immunoprecipitation to study the tyrosine kinase activity of erbB3 against a peptide substrate and its ability to participate in erbB2-erbB3 hetero-interactions and erbB3 homo-interactions
|Steinkamp MP,Low-Nam ST,Yang S,Lidke KA,Lidke DS,Wilson BS||Molecular and cellular biology (34:965)||2014|