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59805
TCRβ (H57-597) Hamster mAb (APC Conjugate)
Antibody Conjugates
Monoclonal Antibody

TCRβ (H57-597) Hamster mAb (APC Conjugate) #59805

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Flow Cytometry Image 1: TCRβ (H57-597) Hamster mAb (APC Conjugate)
Flow cytometric analysis of live C57BL/6 spleen cells using TCRβ (H57-597) Hamster mAb (APC Conjugate) (right) compared to concentration-matched Armenian Hamster Isotype Control (APC Conjugate) (left), co-stained with CD4 (RM4-5) Rat mAb (FITC Conjugate) #96127.
To Purchase # 59805
Cat. # Size Qty. Price
59805S
100 µg $ 160

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Hamster (Armenian) IgG

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to APC and tested in-house for direct flow cytometric analysis in mouse cells.

Product Usage Information

For optimal flow cytometry results, we recommend 0.125 μg of antibody per test.

Application Dilution
Flow Cytometry 1:160

Storage

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 12 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Flow Cytometry, Live Cell Protocol for Directly Conjugated Antibodies

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.

NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised June 2020

Protocol Id: 1504

Specificity / Sensitivity

TCRβ (H57-597) Hamster mAb (APC Conjugate) recognizes endogenous levels of total TCRβ protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Mouse

Source / Purification

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

Background

The αβ T Cell Receptor (TCR) is a heterodimer composed of an alpha chain and a beta chain expressed on the surface of T cells (1,2). It detects peptide antigen presented by major histocompatibility complex (MHC) molecules (1,2). Detection of cognate antigen leads to activation of the TCR signaling pathway through the CD3 signaling chains that associate with the TCR (1). The TCR chains are made up of a constant region and variable regions that get rearranged during T cell development in the thymus (3). There are two TCRβ constant genes, TRBC1 and TRBC2 (3). A population of normal T cells is polyclonal and contains a mix of TRBC1- and TRBC2-expressing T cells, while T cell malignancies are monoclonal and all express the same TCR which either contains TRBC1 or TRBC2 (4). Therefore, it is possible that T cell malignancies could be identified and targeted with reagents or therapies that can distinguish between TRBC1 and TRBC2 (4).
The H57-597 antibody is widely used as a phenotypic marker for T cells expressing the αβ TCR. This antibody does not cross-react with γδ TCR (5).
  1. Kronenberg, M. et al. (1986) Annu Rev Immunol 4, 529-91.
  2. Allison, J.P. et al. (1984) Immunol Rev 81, 145-60.
  3. Saito, H. et al. Nature 309, 757-62.
  4. Maciocia, P.M. et al. (2017) Nat Med 23, 1416-23.
  5. Kubo, R.T. et al. (1989) J Immunol 142, 2736-42.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.
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