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Purified Mouse Anti-Zap70 (pY493)
Purified Mouse Anti-Zap70 (pY493)

Western blot analysis of ZAP70 (pY493).  Lysates from control (left panel) and hydrogen peroxide-treated (right panel) Jurkat cells were probed with purified Mouse anti-ZAP70 (pY493) at concentrations of 8.0 (lanes 1,4), 4.0 (lanes 2,5), and 2.0 µg/ml (Lanes 3,6).  ZAP70 (pY493) is identified as a strong band of 70 kDa in the treated cells.

Purified Mouse Anti-Zap70 (pY493)

ZAP70 staining on tonsil.  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 column) or treated with a phosphatase to eliminate all phosphorylation (right column).  The tissue sections were stained with either purified Mouse anti-ZAP-70 (Cat. No. 610239 or 610240, top row) or purified Mouse anti-ZAP70 (pY493) (bottom row) with Hematoxylin counterstaining.  Original magnification: 20X.

Western blot analysis of ZAP70 (pY493).  Lysates from control (left panel) and hydrogen peroxide-treated (right panel) Jurkat cells were probed with purified Mouse anti-ZAP70 (pY493) at concentrations of 8.0 (lanes 1,4), 4.0 (lanes 2,5), and 2.0 µg/ml (Lanes 3,6).  ZAP70 (pY493) is identified as a strong band of 70 kDa in the treated cells.

ZAP70 staining on tonsil.  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 column) or treated with a phosphatase to eliminate all phosphorylation (right column).  The tissue sections were stained with either purified Mouse anti-ZAP-70 (Cat. No. 610239 or 610240, top row) or purified Mouse anti-ZAP70 (pY493) (bottom row) with Hematoxylin counterstaining.  Original magnification: 20X.

Product Details
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BD Pharmingen™
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Western blot (Routinely Tested), Immunohistochemistry-formalin (antigen retrieval required) (Tested During Development)
70 kDa
0.5 mg/ml
AB_647336
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

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

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
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Antibody Details
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1a/Zap70 (pY493)

The 70-kDa ζ chain-associated protein (ZAP70) is a protein tyrosine kinase (PTK) that associates with the ζ subunit of the T cell antigen receptor (TCR) and undergoes tyrosine phosphorylation following TCR stimulation.  ZAP70 contains two SH2-like domains with the PTK domain located at the C-terminus.  It appears that both ZAP70 and Syk are recruited to the phosphorylated CD3 and ζ subunits after TCR stimulation.  TCR-mediated Lck activity leads to phosphorylation of ZAP70 on Tyrosine 493 (Y493) in the regulatory loop of the kinase domain leading to upregulation of ZAP70 kinase activity.  The significance of ZAP70 activation in mediating TCR signal transduction has been confirmed by showing that ZAP70 activity is absent in an autosomal recessive form of severe combined immunodeficiency (SCID).  This is due to mutations affecting the ZAP70 kinase domain which affect the stability of the protein and TCR signaling.

The 1a/Zap70 (pY493) antibody recognizes ZAP70 phosphorylated at Y493.

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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
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Citations & References
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Development References (3)

  1. Chan AC, Dalton M, Johnson R, et al. Activation of ZAP-70 kinase activity by phosphorylation of tyrosine 493 is required for lmphocyte antigen receptor function. EMBO J. 1995; 14:2499-2508. (Biology). View Reference
  2. Chan AC, Kadlecek TA, Elder ME, et al. ZAP-70 deficiency in an autosomal recessive form of severe combined immunodeficiency. Science. 1994; 264:1599-1601. (Biology).
  3. Mege D, Di Bartolo V, Germain V, Tuosto L, Michel F, Acuto O. Mutation of tyrosines 492/493 in the kinase domain of ZAP-70 affects multiple T-cell receptor signaling pathways. J Biol Chem. 1996; 271:32644-32652. (Biology).
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Please refer to Support Documents for Quality Certificates


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.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.