CD3E (activation epitope) Mouse Monoclonal Antibody [Clone ID: APA1/1]

Specifications

Product Data
Clone Name	APA1/1
Applications	FC, IF, IHC, IP, WB
Recommend Dilution	Flow Cytometry. Sample preparation: At the end of stimulation of T cells, perform staining of surface markers (if required) in PBS + 0.1% BSA for 20 min. on ice. Wash with PBS and fix with 2% formaldehyde, 30 min on ice. Wash with PBS and incubate in PBS + 0,1% saponine, 5 min. RT. Incubate the cells in PBS + 1% BSA + 0.03% saponine, 15 min. on ice. Incubate with fluorescence-labeled APA1/1 antibody (1-5 µg/ml) in PBS + 1% BSA + 0.03% saponine in dark, 20 min. RT. Wash with PBS + 1% BSA + 0.03% saponine, resuspend in PBS. Positive control: Human T cells stimulated with anti-CD3 (MEM-57) antibody (1 µg/ml). Immunoprecipitation. Western blot. Immunohistochemistry on frozen sections. Immunocflourescence. Application note: Fixed and permeabilised cells. The antibody can distinguish TCR-stimulated from non-stimulated cells.
Reactivity	Human, Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	Purified human CD3 proteins isolated from thymus
Specificity	The antibody recognizes an activation-dependent intracellular epitope of CD3 epsilon. Exposure of the epitope precedes CD3 phosphorylation and recruitment and activation of ZAP70, which initiates the signaling cascade produced by T-cell activation. APA1/1 provides the earliest known marker for TCR-mediated T cell activation.
Formulation	PBS, pH 7.4 State: Aff - Purified State: Liquid Ig fraction Preservative: 15 mM sodium azide
Concentration	lot specific
Purification	Protein A affinity chromatography (> 95% pure by SDS-PAGE)
Conjugation	Unconjugated
Storage Condition	Store at 2 - 8 °C for up to one month or (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Gene Name	CD3e molecule
Database Link	Entrez Gene 12501 MouseEntrez Gene 916 Human UniProt ID P07766
Background	CD3 complex is crucial in transducing antigen-recognition signals into the cytoplasm of T cells and in regulating the cell surface expression of the TCR complex. T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3 gamma, CD3 delta, CD3 epsilon and CD3 zeta. These CD3 subunits are structurally related members of the immunoglobulins super family encoded by closely linked genes on human chromosome 11. The CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine-based motif present in a single copy in the CD3 subunits. CD3 may play a role in TCR-induced growth arrest, cell survival and proliferation.
Synonyms	T3/Leu-4
Reference Data
Protein Families	Druggable Genome, Transmembrane
Protein Pathways	Hematopoietic cell lineage, Primary immunodeficiency, T cell receptor signaling pathway

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