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9382
T Cell Signaling Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

T Cell Signaling Antibody Sampler Kit #9382

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T Cell Signaling Antibody Sampler Kit: Image 1
Western blot analysis of extracts from Jurkat cells treated with hydrogen peroxide (2mM for 2 minutes) or with lambda phosphatase and extracts from Ramos cells treated with anti-human IgM (12 micrograms/ml for 2 minutes) using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb.
T Cell Signaling Antibody Sampler Kit: Image 2
Confocal immunofluorescent analysis of Ramos cells, human IgM treated (left) or untreated (right), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.
T Cell Signaling Antibody Sampler Kit: Image 3
Flow cytometric analysis of Ramos cells, untreated (blue) or treated with anti-IgM (green), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.
T Cell Signaling Antibody Sampler Kit: Image 4
Western blot analysis of extracts from Jurkat cells (starved for 16 hours) treated with calf intestinal alkaline phosphatase (CIP) or H2O2 (2 mM), using Phospho-Lck (Tyr505) Antibody (upper) or control Lck Antibody #2752 (lower).
T Cell Signaling Antibody Sampler Kit: Image 5
Western blot analysis of Jurkat cell extracts using Lck (73A5) Rabbit mAb.
T Cell Signaling Antibody Sampler Kit: Image 6
Confocal immunofluorescent analysis of a Jurkat cell nucleus using Lck (73A5) Rabbit mAb (red) and CD3 antibody (green). Blue=DRAQ5® #4084 fluorescent DNA dye.
T Cell Signaling Antibody Sampler Kit: Image 7
Flow cytometric analysis of Jurkat cells, using Lck (73A5) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
T Cell Signaling Antibody Sampler Kit: Image 8
Western blot analysis of extracts from Jurkat cells using Zap-70 (D1C10E) XP® Rabbit mAb.
T Cell Signaling Antibody Sampler Kit: Image 9
Immunoprecipitation of Zap-70 from Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Zap-70 (D1C10E) XP® Rabbit mAb. Western blot analysis was performed using Zap-70 (D1C10E) XP® Rabbit mAb. Mouse Anti-rabbit (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
T Cell Signaling Antibody Sampler Kit: Image 10
Confocal immunofluorescent analysis of Jurkat cells (left) and Ramos cells (right) using Zap-70 (D1C10E) XP® Rabbit mAb #3165 (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
T Cell Signaling Antibody Sampler Kit: Image 11
Western blot analysis of SDS extracts from untreated or anti-CD3 treated (10 µg/ml for 2 minutes) human Jurkat cells after overnight serum starvation using Phospho-LAT (Tyr220) Antibody.
T Cell Signaling Antibody Sampler Kit: Image 12
Western blot analysis of extracts from Jurkat and BW5147cells using SLP-76 Antibody.
T Cell Signaling Antibody Sampler Kit: Image 13
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.
T Cell Signaling Antibody Sampler Kit: Image 14
Western blot analysis of extracts from Jurkat cells, untreated or treated with lambda phosphatase, using LAT Antibody.
T Cell Signaling Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded Jurkat cells, using LAT Antibody in the presence of control peptide (left) or antigen-specific peptide (right).
T Cell Signaling Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded human tonsil, showing membrane localization, using LAT Antibody.
T Cell Signaling Antibody Sampler Kit: Image 17
Flow cytometric analysis of Jurkat cells, using LAT Antibody (blue) compared to a nonspecific negative control antibody (red).
Inquiry Info.# 9382

Product Description

The T Cell Signaling Antibody Sampler Kit contains primary and secondary antibodies to perform four Western blots with each antibody.

Specificity / Sensitivity

Each antibody in the T Cell Signaling Antibody Sampler Kit recognizes only its specific target and does not cross-react with other family members with the following exceptions: Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb which also recognizes Syk when phosphorylated at Tyr352; and Phospho-Lck (Tyr505) Antibody which may cross-react with certain phosphorylated Src family members.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the amino terminus of human Zap-70; Tyr319 of human Zap-70; and Tyr505 of human Lck. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human SLP-76; residues surrounding Tyr191 of human LAT; the central region of human LAT; and residues surrounding Tyr505 of human Lck. Antibodies are purified by protein A and peptide affinity chromatography.

Background

The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8). LAT, a transmembrane adaptor protein expressed in T, NK and mast cells, is an important mediator for T cell receptor (TCR) signaling (9). Upon TCR engagement, activated Zap-70 phosphorylates LAT at multiple conserved tyrosine residues within SH2 binding motifs, exposing these motifs as the docking sites for downstream signaling targets (10,11). The phosphorylation of LAT at Tyr171 and 191 enables the binding of Grb2, Gads/SLP-76, PLCgamma1 and PI3 kinase through their SH2 domain and translocates them to the membrane. This process eventually leads to activation of the corresponding signaling pathways (11-12).
  1. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
  2. Iwashima, M. et al. (1994) Science 263, 1136-9.
  3. Neumeister, E.N. et al. (1995) Mol Cell Biol 15, 3171-8.
  4. Chan, A.C. et al. (1995) EMBO J 14, 2499-508.
  5. Williams, B.L. et al. (1999) EMBO J 18, 1832-44.
  6. Di Bartolo, V. et al. (1999) J Biol Chem 274, 6285-94.
  7. Wiestner, A. et al. (2003) Blood 101, 4944-51.
  8. Crespo, M. et al. (2003) N Engl J Med 348, 1764-75.
  9. Wonerow, P. and Watson, S.P. (2001) Oncogene 20, 6273-83.
  10. Zhang, W. et al. (1998) Cell 92, 83-92.
  11. Paz, P.E. et al. (2001) Biochem J 356, 461-71.
  12. Zhang, W. et al. (2000) J Biol Chem 275, 23355-61.

Pathways & Proteins

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Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.