Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

EGF Receptor vIII (D6T2Q) XP® Rabbit mAb #64952

EGF Receptor   EGFR   EGFR (Val30-Arg297 del) mutation   EGFR mutation   EGFR viii  

No. Size Price
64952S 100 µl ( 200 sections ) ¥3,580.00 现货查询 购买询价
64952T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
64952 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
IHC-P 1:200 Human, Endogenous (IHC-P), Transfected (IF) 130 Rabbit IgG
IF-IC 1:3200

Species cross-reactivity is determined by western blot.

Applications Key: IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

EGF Receptor vIII (D6T2Q) XP® Rabbit mAb recognizes endogenous and transfected levels of EGF Receptor vIII protein by immunohistochemistry and immunofluorescence, respectively.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human EGF Receptor vIII protein.

IF-IC

IF-IC

Confocal immunofluorescent analysis of COS-7 cells, mock-transfected (left), or transiently transfected with EGF Receptor vIII mutation construct (center), and mock-transfected A549 cells (right), using EGF Receptor vIII (D6T2Q) XP® Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087. Note the lack of EGFR vIII staining in A549 cells expressing wild-type human EGFR.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human glioblastoma using EGF Receptor vIII (D6T2Q) XP® Rabbit mAb in the presence of control peptide (left) and antigen-specific peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human glioblastoma using EGF Receptor vIII (D6T2Q) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded EGF Receptor vIII transfected 293T cell pellets (left) and HCC827 (EGFR positive) xenograft (middle and right) using EGF Receptor vIII (D6T2Q) XP® Rabbit mAb (left and middle) and total EGF Receptor (D38B1) XP® Rabbit mAb #4267 (right). Note the lack of EGFR vIII staining in the HCC827 xenograft that is strongly positive for total EGFR.

Background

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). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

EGFR variant III (EGFRvIII) is a truncated, constitutively active mutant form of EGFR that results from an in-frame deletion of exons 2-7 (11,12). EGFRvIII is expressed in various cancers, most notably glioblastoma, where it is expressed at a frequency of 25-30%. Although complicated by the fact that it is often co-expressed with amplified EGFR, EGFRvIII is a potential therapeutic target (13).

  1. Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
  2. Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
  3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
  4. Hubbard, S.R. et al. (1994) Nature 372, 746-54.
  5. Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
  6. Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
  7. Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
  8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
  9. Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
  10. Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.
  11. Sugawa, N. et al. (1990) Proc Natl Acad Sci U S A 87, 8602-6.
  12. Wong, A.J. et al. (1992) Proc Natl Acad Sci U S A 89, 2965-9.
  13. Padfield, E. et al. (2015) Front Oncol 5, 5.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SignalStain is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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