Description
The T cell receptor (TCR), expressed by thymocytes and T lymphocytes, is a multi-component cell-surface complex responsible for recognizing antigen in the context of MHC molecules. The antigen-specific binding component of the TCR, Ti, is a heterodimer of the variable Ig-like subunits α and β or γ and δ. Ti is non-covalently associated with an invariant set of molecules referred to as the CD3 polypeptides, γ, δ, ε, and ζ. The CD3 ζ polypeptide (CD3ζ) was named CD247 at the 7th Human Leukocyte Differentiation Antigens Workshop. CD3 appears early in thymocyte differentiation and remains expressed on all mature T lymphocytes. After antigen recognition by the TCR, CD3ζ is the primary intracellular signal transducing subunit. It contains three ITAMs (Immunoreceptor Tyrosine-based Activation Motifs), each of which contains a pair of tyrosine residues that are phosphorylated by Lck and Fyn and are required for signal propagation. The molecular weight of CD3ζ is 16 kDa, and it is also observed as 32-kDa homodimers or as heterodimers with the γ chain of Fc receptors. Upon phosphorylation, the CD3ζ monomer undergoes an apparent shift in electrophoretic mobility up to 21 kDa. The K25-407.69 monoclonal antibody recognizes the phosphorylated tyrosine 142 (pY142) in the third ITAM domain of human CD3ζ (CD247).
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
3. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
5. 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.
6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
