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9923
Phospho-HER2/ErbB2 Antibody Sampler Kit

Phospho-HER2/ErbB2 Antibody Sampler Kit #9923

Western Blotting Image 1

Western blot analysis of activated recombinant HER2/ErbB2 and HER1/EGFR tyrosine kinase proteins, using Phospho-HER2/ErbB2 (Tyr1248) (upper), EGF Receptor (D38B1) XP® Rabbit mAb #4267 (middle) or HER2/ErbB2 (29D8) Rabbit mAb #2165 (lower). Phospho-HER2/ErbB2 (Tyr1248) specifically detects phosphorylated HER2/ErbB2 but not phosphorylated HER1/EGFR.

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Western Blotting Image 2

Western blot analysis of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb (upper) or Phospho-Tyrosine 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.

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Western Blotting Image 3

Western blot analysis of extracts from SK-BR-3 and MCF7 cells using HER2/ErbB2 (D8F12) XP® Rabbit mAb.

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Western Blotting Image 4

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Western Blotting Image 5

Western blot analysis of T47D and MCF7 cells untreated or treated with Neuregulin (100 ng/ml) for 5 minutes, using Phospho-HER2/ErbB2 (Tyr1248) Antibody (upper) or HER2/ErbB2 (D8F12) XP® Rabbit mAb #4290 (lower).

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IHC-P (paraffin) Image 6

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing membrane localization, using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb.

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IHC-P (paraffin) Image 7

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HER2/ErbB2 (D8F12) XP® Rabbit mAb.

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IHC-P (paraffin) Image 8

Immunohistochemical analysis of paraffin-embedded SkBr3 cell pellets untreated (left) EGF-treated (right) either untreated (top) or lambda-phosphatase-treated (bottom), using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb.

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IHC-P (paraffin) Image 9

Immunohistochemical analysis of paraffin-embedded SK-BR-3 (Her2 high, left) and MCF7 cell pellets (Her2 low, right) using HER2/ErbB2 (D8F12) XP® Rabbit mAb.

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human renal adenocarcinoma, using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb in the presence of control peptide (left) or Phospho-HER2/ErbB2 (Tyr1221/1222) Blocking Peptide #1254 (right).

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IHC-P (paraffin) Image 11

Immunohistochemical analysis of paraffin-embedded human NCI-H358 xenograft, using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb.

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IHC-F (frozen) Image 12

Immunohistochemical analysis of frozen HCC827 xenograft, using Phospho-Her2/ErbB2 (Tyr1221/1222)(6B12) Rabbit mAb.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-HER2/ErbB2 (Tyr1248) Antibody 2247 20 µl
  • WB
H M 185 Rabbit 
Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb 2243 20 µl
  • WB
  • IHC
H M 185 Rabbit IgG
HER2/ErbB2 (D8F12) XP® Rabbit mAb 4290 20 µl
  • WB
  • IHC
H M 185 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Phospho-HER2/ErbB2 Antibody Sampler Kit provides an economical means to evaluate the activation status of HER2/ErbB2, including the phosphorylation of Tyr1248 and Tyr1221/1222. The control HER2/ErbB2 antibody is also included. The kit contains enough primary antibodies to perform two Western blot experiments per primary antibody.

Each phospho-HER2/ErbB2 antibody recognizes only the phosphorylated form of HER2/ErbB2 at the indicated sites. The control HER2/ErbB2 receptor antibody recognizes both phosphorylated and nonphosphorylated forms of HER2/ErbB2.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1248. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding tyrosines 1221/1222 and residues near the amino terminus of human ErbB2 protein.

The ErbB2 (HER2) proto-oncogene encodes a 185 kDa transmembrane, receptor-like glycoprotein with intrinsic tyrosine kinase activity (1). 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 (2). Amplification of the ErbB2 gene and overexpression of its product are detected in almost 40% of human breast cancers (3). Binding of the c-Cbl ubiquitin ligase to ErbB2 at Tyr1112 leads to ErbB2 poly-ubiquitination and enhances degradation of this kinase (4). ErbB2 is a key therapeutic target in the treatment of breast cancer and other carcinomas and targeting the regulation of ErbB2 degradation by the c-Cbl-regulated proteolytic pathway is one potential therapeutic strategy. Phosphorylation of the kinase domain residue Tyr877 of ErbB2 (homologous to Tyr416 of pp60c-Src) may be involved in regulating ErbB2 biological activity. 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 (1,5).

  1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
  2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
  3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
  4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
  5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.
Entrez-Gene Id
2064
Swiss-Prot Acc.
P04626
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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