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Anti-erbB-2 Antibody, clone N3/D10 (Cytoplasmic Domain)
Recommended Replacement for: 05-1130 is a recommended replacement for CBL755
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Species Reactivity Key Applications Host Format Antibody Type
H, M, R WB, ICC, IH(P), FC Mouse Affinity Purified Monoclonal Antibody
Description:
Anti-erbB-2 Antibody, clone N3/D10 (Cytoplasmic Domain)
Replaces:
05-1130 is a recommended replacement for CBL755
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Specificity:
The antibody reacts with c-erbB-2 which is a membrane associated oncoprotein that has considerable sequence homology with EGFR. Membrane preparations were made from SK-BR-3 mammary adenocarcinoma and A431 epidermoid carcinoma cell lines. SK-BR-3 expresses high levels of c-erbB-2 whilst A431 expresses high levels of EGFR.
The antibody has been used to detect c-erbB-2 in paraffin-embedded material from breast tumours.
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Molecular Weight:
~138 kDa
Epitope:
a.a. 1238-1255, Cytoplasmic Domain
Immunogen:
Synthetic peptide:
Cys-Lys-Gly-Thr-Pro-Thr-Ala-Glu-Asp-Pro-Glu-Tyr-Leu-Gly-Leu-Asp-Val-Pro-Val (amino acids 1238-1255) selected from the cytoplasmic domain of human c-erbB-2 protein.
Clone:
N3/D10
Isotype:
IgG1
Background Information:
v-ErbB was identified as the viral counterpart of the EGF Receptor, and its relatives have adopted this nomenclature (or the HER nomenclature for human proteins). These proteins are activated by Heregulin, Neuregulin and Amphiregulin. ErbB-2/HER-2, also known as neu, is amplified in large numbers of highly aggressive breast cancers, but all the family members are capable of heterodimerization, and together activate downstream signaling pathways. ErbB-3 has a natural "substitution" at the codon which is an invariant Lys residue at the ATP-binding site of all protein kinases. It is therefore inactive as a kinase, and can only become phosphorylated by heterodimerization with another erbB-family member. All members of this family, when Tyr-phosphorylated, can nucleate the formation of signaling complexes through adapter proteins.
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Species Reactivity:
Human
Mouse
Rat
Species Reactivity Note:
Human, mouse and rat.
Application Notes:
Immunocytochemistry: A previous lot of this antibody was used in immunocytochemistry on frozen sections.
Flow Cytometry: A previous lot of this antibody was used with FC.
Optimal working dilutions must be determined by the end user.
Immunohistochemistry(paraffin): Representative testing from a previous lot.
Optimal Staining of erbB-2/HER-2 Monoclonal Antibody: Breast Cancer
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Control:
A431 Cell Lysate
Quality Assurance:
Routinely evaluated by Western Blot on A431 lysates.
Western Blot Analysis: 1:500 dilution of this lot detected C-ERBB-2 on 10 μg of A431 lysates.
Purification Method:
Protein A affinity chromatography
Presentation:
Purified mouse monoclonal IgG1 is presented in phosphate buffered saline containing 10mM sodium azide and 1mg/ml bovine serum albumin. We recommend that each laboratory determine an optimum working titre for use in its particular application.
Storage Conditions:
For use within 1 month of purchase store at 2-8°C, for long term storage aliquot antibody into small volumes and store at -20°C for up to 1 year from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoi |