Product Pathways - Tyrosine Kinase / Adaptors
HER/ErbB Family Antibody Sampler Kit #8339
|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|EGF Receptor (D38B1) XP® Rabbit mAb #4267||40 µl||W IP IHC-P IF-IC F||H M Mk||175||Rabbit IgG|
|HER2/ErbB2 (D8F12) XP® Rabbit mAb #4290||40 µl||W IHC-P||H (M) (R)||185||Rabbit IgG|
|HER3/ErbB3 (1B2E) Rabbit mAb #4754||40 µl||W IP||H (M) (R)||185||Rabbit IgG|
|HER4/ErbB4 (111B2) Rabbit mAb #4795||40 µl||W IP||H M (R)||180||Rabbit IgG|
|Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777||40 µl||W IHC-P IF-IC F||H M R Mk||175||Rabbit IgG|
|Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb #2243||40 µl||W IHC-P IHC-F||H||185||Rabbit IgG|
|Phospho-HER3/ErbB3 (Tyr1222) (50C2) Rabbit mAb #4784||40 µl||W||H||185||Rabbit IgG|
|Phospho-HER4/ErbB4 (Tyr1284) (21A9) Rabbit mAb #4757||40 µl||W||H (M) (R)||180||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Each antibody in the HER/ErbB Family Antibody Sampler Kit recognizes only its specific target. The antibodies do not cross react with other HER/ErbB family members.
Western blot analysis of extracts from various cell lines expressing different activated tyrosine kinase proteins using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb #2243 (upper) or Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (lower). Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb specifically detects phosphorylated HER2/ErbB2 but not other phosphorylated tyrosine kinases.
Western blot analysis of extracts of BxPC-3 cells, untreated or EGF-stimulated, using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 (upper) or EGF Receptor Antibody #2232 (lower).
Western blot analysis of extracts from various cell lines using EGF Receptor (D38B1) XP® Rabbit mAb #4267.
Western blot analysis of extracts from SK-BR-3 and MCF7 cells using HER2/ErbB2 (D8F12) XP® Rabbit mAb #4290.
Western blot analysis of extracts from MCF7 and T47D cell using HER3/ErbB3 (1B2E) Rabbit mAb #4754.
Western blot analysis of extracts from various cell lines expressing different activated protein tyrosine kinases using Phospho-HER4/ErbB4 (Tyr1284) (21A9) Rabbit mAb #4757 or Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (lower).
Western blot analysis of extracts from MCF7 and T47D cells, untreated or stimulated with Neuregulin (NRG; 100 ng/ml, 15 min), using Phospho-HER3/ErbB3 (Tyr1222) (50C2) Rabbit mAb #4784 (upper) or HER3/ErbB3 (1B2E) Rabbit mAb #4754 (lower).
The HER/ErbB Family Antibody Sampler Kit provides an economical means to evaluate the HER/ErbB Family, including the phosphorylation of EGFR, HER2/ErbB2, HER3/ErbB3, and HER4/ErbB4. The control antibodies to each family member are also included. The kit contains enough antibody to perform four western blot experiments with each primary antibody.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a fusion protein containing the cytoplasmic domain of human EGF receptor, a synthetic peptide corresponding to residues near the amino terminus of human ErbB2 protein, Tyr867 of human ErbB3 protein, or the carboxy-terminus of human ErbB4 protein. Activation state monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1068 of human EGF receptor protein, tyrosines 1221/1222 of human ErbB2 protein, Tyr1222 of human ErbB3 protein, or Tyr1284 of human ErbB4 protein.
The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (3).The ErbB2 (HER2) proto-oncogene encodes a 185 kDa transmembrane, receptor-like glycoprotein with intrinsic tyrosine kinase activity (4). While ErbB2 lacks an identified ligand, ErbB2 kinase activity can be activated in the absence of a ligand when overexpressed and through heteromeric associations with other ErbB family members (5). Amplification of the ErbB2 gene and overexpression of its product are detected in almost 40% of human breast cancers (6). The major autophosphorylation sites in ErbB2 are Tyr1248 and Tyr1221/1222; phosphorylation of these sites couples ErbB2 to the Ras-Raf-MAP kinase signal transduction pathway (4,7).HER3/ErbB3 is a member of the ErbB receptor protein tyrosine kinase family, but lacks tyrosine kinase activity. Tyrosine phosphorylation of ErbB3 depends on its association with other ErbB tyrosine kinases. Upon ligand binding, heterodimers form between ErbB3 and other ErbB proteins, and ErbB3 is phosphorylated on tyrosine residues by the activated ErbB kinase (8,9). There are at least 9 potential tyrosine phosphorylation sites in the carboxy-terminal tail of ErbB3. These sites serve as consensus binding sites for signal transducing proteins, including Src family members, GRB2, and the p85 subunit of PI3 kinase, which mediate ErbB-downstream signaling (10). Both Tyr1222 and Tyr1289 of ErbB3 reside within a YXXM motif and participate in signaling to PI3 kinase (11).HER4/ErbB4, like other family members, has four ectodomains, a single transmembrane domain, and a cytoplasmic tail containing the active tyrosine kinase domain (12). By binding to neuregulins and/or EGF family ligands, ErbB4 forms either a homodimer or heterodimer with other ErbB family members, which results in receptor activation and signaling (12). ErbB4 is ubiquitously expressed with the highest expression occurring in the brain and heart. The expression of ErbB4 in breast cancer, pediatric brain cancer, and other types of carcinomas has been reported, suggesting that ErbB4 expression is involved in both normal tissue development and carcinogenesis (12).
- Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
- Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
- Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
- Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
- Qian, X. et al. (1994) Proc Natl Acad Sci U S A 91, 1500-4.
- Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
- Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.
- Yarden, Y. and Sliwkowski, M.X. (2001) Nat Rev Mol Cell Biol 2, 127-37.
- Guy, P.M. et al. (1994) Proc Natl Acad Sci U S A 91, 8132-6.
- Songyang, Z. et al. (1993) Cell 72, 767-78.
- Kim, H.H. et al. (1994) J Biol Chem 269, 24747-55.
- Carpenter, G. (2003) Exp Cell Res 284, 66-77.
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For Research Use Only. Not For Use In Diagnostic Procedures.