Alexa Fluor® 647 Mouse Anti-Human CD3 ζ (CD247)
Clone 6B10.2 (RUO)
- Brand BD Pharmingen™
- Alternative Name CD247; CD3-ZETA; CD3Z; TCR zeta chain; TCRZ; T3Z
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Human TCR zeta Peptide
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 6B10.2 monoclonal antibody specifically recognizes CD3 zeta (CD3Z or CD3ζ), which is also known as either CD247 or the invariant TCR zeta chain (TCRZ, T3Z, or TCRζ). CD3ζ (CD247) is a ~16 kDa single-pass, type I transmembrane protein that belongs to the CD3Z/FCER1G family. CD3ζ (CD247) has a short extracellular domain of nine amino acids and a cytoplasmic domain that contains three immunoreceptor tyrosine-based activation motifs (ITAMs). Disulfide-linked CD3ζ homodimers usually associate with other transmembrane CD3 protein subunits (CD3δ, CD3ε, and CD3γ) to form the CD3 signaling complex which is linked to the clonotypic T cell receptor (TCR) for antigen, αβ TCR or γδ TCR. CD3ζ (CD247) is also expressed by natural killer (NK) cells where it can function as a signaling subunit associated with CD16a (FcγRIIIa), a receptor for IgG complexes. Human CD16a can couple with either CD3ζ or FcεRIγ homodimers, or CD3ζ-FcεRIγ heterodimers that serve to transduce IgG-bound CD16a-mediated signals intracellularly. Low or absent CD3ζ (CD247) expression is reportedly associated with impaired immune responses leading to some chronic infections or autoimmune diseases, and certain cancers. The 6B10.2 antibody is directed against the cytoplasmic domain of human CD3ζ (CD247).
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.