Rabbit ERBB2 / HER2 Polyclonal Antibody | anti-ERBB2 antibody
Rabbit Polyclonal (IgG) to Human ERBB2 / HER2
Predicted Reactivity: Bat, Bovine
Predicted Reactivity: Bat, Bovine
IHC-paraffin: 5 ug/ml
Western Blot: 0.5-2ug/ml
Tissue Preparation:
Formalin fixation and embedding in paraffin wax.
Tissue Sectioning:
Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C.
Deparaffinization:
Wash dry slides in 3 changes of xylene - 5 minutes each @ RT
Rehydration:
Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT
Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT
Wash slides in 1 change of 80% alcohol - 3 minutes @ RT
Rinse slides in gentle running distilled water - 5 minutes @ RT
Antigen retrieval:
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) -1 minute @ RT
Immunostaining:
(Do not allow tissues to dry at any time during the staining procedure)
Apply a universal protein block - 20 minutes @ RT
Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT
Rinse slides in 1 X TBST - 1 minute @ RT
Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase streptavidin - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT
Wash slides in distilled water - 1 minute @ RT
Dehydrate:
(This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB)
Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT
Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT
Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT
Wash slides in 3 changes of xylene - 1 minute each @ RT
Apply coverslip
NCBI and Uniprot Product Information
NCBI Description
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. [provided by RefSeq, Jul 2008]
Uniprot Description
HER2: a proto-oncogenic receptor tyrosine kinase of the EGFR family. Essential component of a neuregulin-receptor complex, although neuregulins do not interact with it alone. Not activated by EGF, TGF- alpha and amphiregulin. Amplified in breast cancer. Overexpression induces constitutive activity, and the gene is amplified or overexpressed in up to 30% of breast cancers, correlating with poor survival. The antibody Herceptin is approved for treatment of metastatic breast cancer with HER2 amplification/overexpression. Somatic mutations seen in 4% of lung cancers and also in breast, gastric, ovarian cancer and glioblastoma. One SNP shows predisposition to breast and gastric cancer. Inhibitors: Herceptin, lapatinib, PKI-166, EKB-569, CI-1033.
Protein type: Oncoprotein; Protein kinase, tyrosine (receptor); EC 2.7.10.1; Protein kinase, TK; Kinase, protein; Membrane protein, integral; TK group; EGFR family
Chromosomal Location of Human Ortholog: 17q12
Cellular Component: perinuclear region of cytoplasm; basolateral plasma membrane; apical plasma membrane; cytoplasm; integral to membrane; plasma membrane; endosome membrane; cytoplasmic vesicle; nucleus; receptor complex
Molecular Function: protein C-terminus binding; protein dimerization activity; identical protein binding; receptor signaling protein tyrosine kinase activity; transmembrane receptor protein tyrosine kinase activity; protein phosphatase binding; protein binding; transmembrane receptor activity; ErbB-3 class receptor binding; protein heterodimerization activity; protein-tyrosine kinase activity; growth factor binding; ATP binding
Biological Process: myelination; axon guidance; phosphoinositide 3-kinase cascade; positive regulation of cell adhesion; peptidyl-tyrosine phosphorylation; nerve growth factor receptor signaling pathway; positive regulation of translation; wound healing; heart development; protein amino acid autophosphorylation; motor axon guidance; signal transduction; protein amino acid phosphorylation; enzyme linked receptor protein signaling pathway; positive regulation of transcription from RNA polymerase I promoter; positive regulation of MAP kinase activity; cell surface receptor linked signal transduction; oligodendrocyte differentiation; neuromuscular junction development; epidermal growth factor receptor signaling pathway; negative regulation of immature T cell proliferation in the thymus; regulation of microtubule-based process; fibroblast growth factor receptor signaling pathway; phosphoinositide-mediated signaling; transcription, DNA-dependent; regulation of angiogenesis; positive regulation of cell growth; peripheral nervous system development; cell proliferation; positive regulation of transcription from RNA polymerase III promoter; innate immune response; positive regulation of protein amino acid phosphorylation; transmembrane receptor protein tyrosine kinase signaling pathway; positive regulation of epithelial cell proliferation
Disease: Gastric Cancer; Lung Cancer; Glioma Susceptibility 1
Research Articles on ERBB2
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Product Notes
The ERBB2 erbb2 (Catalog #AAA241896) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Rabbit Polyclonal (IgG) to Human ERBB2 / HER2 reacts with Human, Monkey, Mouse, Rat, Dog, Hamster, Horse, Pig Predicted Reactivity: Bat, Bovine and may cross-react with other species as described in the data sheet. AAA Biotech's ERBB2 / HER2 can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC - Paraffin; IHC - Frozen), Western Blot (WB), Immunoprecipitation (IP). Immunohistochemistry: MBS241896 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for MBS241896 was determined to be 5 ug/ml. Western Blot: 0.5-2 ug/ml detects c-erbB-2 in RIPA lysates from A431, MDA, MCF-7, LNCap, and c-erbB-2 transfected 3T3/NIH cells. Immunoprecipitation: 4 ug immunoprecipitated c-erbB-2 from 500 ug of A431, LNCap, and c-erbB- 2 transfected 3T3/NIH RIPA lysate. IHC-paraffin: 5 ug/ml Western Blot: 0.5-2ug/ml. Researchers should empirically determine the suitability of the ERBB2 erbb2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "ERBB2 / HER2, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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