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Purified Mouse anti-EGF Receptor (pY1173)
Purified Mouse anti-EGF Receptor (pY1173)

Western blot analysis of EGF Receptor (pY1173) in human epidermis (left figure).  Lysates from control (left panel) and EGF-treated (Cat. No. 354052, right panel) human A-431 epidermoid carcinoma were probed with the  purified mouse anti-EGF Receptor (pY1173) monoclonal antibody at concentrations of 0.4 µg/mL, 0.2 µg/mL, and 0.1 µg/mL (lanes 1, 2, and 3, respectively).  EGF Receptor (pY1173) is identified as a band of 175 kDa in the treated cells.  Immunohistochemical analysis for EGF Receptor (pY1173) staining on tonsil (right figure).  Fresh human tonsil was incubated in 5 mM Pervanadate solution for 2 hours, then fixed in formalin and processed.  Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the sections were either left untreated (left panel) or treated with a phosphatase to eliminate all phosphorylation (right panel).  The tissue sections were stained with purified Mouse anti-EGF Receptor (pY1173) with Hematoxylin counterstaining.  No staining was seen on sections of unstimulated tonsil (data not shown).  Original magnification: 20X.

Western blot analysis of EGF Receptor (pY1173) in human epidermis (left figure).  Lysates from control (left panel) and EGF-treated (Cat. No. 354052, right panel) human A-431 epidermoid carcinoma were probed with the  purified mouse anti-EGF Receptor (pY1173) monoclonal antibody at concentrations of 0.4 µg/mL, 0.2 µg/mL, and 0.1 µg/mL (lanes 1, 2, and 3, respectively).  EGF Receptor (pY1173) is identified as a band of 175 kDa in the treated cells.  Immunohistochemical analysis for EGF Receptor (pY1173) staining on tonsil (right figure).  Fresh human tonsil was incubated in 5 mM Pervanadate solution for 2 hours, then fixed in formalin and processed.  Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the sections were either left untreated (left panel) or treated with a phosphatase to eliminate all phosphorylation (right panel).  The tissue sections were stained with purified Mouse anti-EGF Receptor (pY1173) with Hematoxylin counterstaining.  No staining was seen on sections of unstimulated tonsil (data not shown).  Original magnification: 20X.

Product Details
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BD Pharmingen™
Human (QC Testing), Mouse, Rat, Dog (Reported)
Mouse IgG1, κ
Phosphorylated Human EGF Receptor Peptide
Western blot (Routinely Tested), ELISA, Immunohistochemistry-formalin (antigen retrieval required), Immunoprecipitation (Reported)
175 kDa
0.5 mg/ml
AB_647272
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558382 Rev. 3
Antibody Details
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9H2

Epidermal Growth Factor (EGF) elicits a variety of cellular responses that are initiated by EGF Receptor (EGFR) binding and activation of intrinsic tyrosine kinase activity.  EGFR, also known as ErbB1 or HER1, is a member of the ErbB class of receptor protein tyrosine kinases.  It has an extracellular ligand-binding domain, a single transmembrane region, and a cytoplasmic region containing a protein tyrosine kinase domain and a c-terminal regulatory domain with many phosphorylation sites.  Following ligand binding, EGFR forms homodimers and heterodimers with ErbB2.  Specific C-terminal tyrosine residues are then autophosphorylated and, in turn, bind to adaptor proteins, kinases, or protein tyrosine phosphatases.  Specifically, phosphorylated tyrosine 1173 (Y1173) interacts with Shc, SHP1, and PLCγ, which mediate downstream signaling cascades and negative feedback regulation of EGFR activation.  Inappropriate expression or mutations of EGFR and/or deregulation of its signaling pathways are associated with many types of cancer, making EGFR a promising target for cancer therapies.

The 9H2 monoclonal antibody recognizes the phosphorylated Y1173 in the regulatory domain of human EGFR.

558382 Rev. 3
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
558382 Rev.3
Citations & References
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Development References (3)

  1. Hristozova T, Konschak R, Budach V, Tinhofer I. A simple multicolor flow cytometry protocol for detection and molecular characterization of circulating tumor cells in epithelial cancers. Cytometry A. 2012; 81A(6):489-495. (Clone-specific: Flow cytometry). View Reference
  2. Mendelsohn J, Baselga J. Status of epidermal growth factor receptor antagonists in the biology and treatment of cancer. J Clin Oncol. 2003; 21(14):2787-2799. (Biology).
  3. Olayioye MA, Neve RM, Lane HA, Hynes NE. The ErbB signaling network: receptor heterodimerization in development and cancer. EMBO J. 2000; 19(13):3159-3167. (Biology).
558382 Rev. 3

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

Non-IVD products are For Research Use Only. Not for use in diagnostic or therapeutic procedures.

 

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