|Flow Cytometry (Flow)||5 µL (0.25 µg)/test|
|Immunoprecipitation (IP)||See 1 publications below|
|Miscellaneous PubMed (MISC)||See 1 publications below|
|Western Blot (WB)||See 1 publications below|
|Tested Species reactivity||Human, Rhesus monkey|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG2a, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin, 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage conditions||4° C, store in dark, DO NOT FREEZE!|
Description: The 14D3 monoclonal antibody reacts with human CD152, also known as cytotoxic T lymphocyte antigen-4 (CTLA-4). CTLA-4, a protein with structural similarities to CD28, is expressed on activated T cells (activated B cells may also express CTLA-4) and binds the B7 family members, CD80 (B7-1) and CD86 (B7-2), with higher affinity than CD28 does. CTLA-4 and CD28 appear to deliver opposing signals to T cells: while CD28 is a potent costimulator, CTLA-4 restricts the progression of T cells to an activated state by inhibiting IL-2 secretion and cellular proliferation. The cytoplasmic portion of CTLA-4 contains ER retention motifs, resulting in intracellular localization of a large proportion of newly synthesized CTLA-4 in response to TCR signaling.
The 14D3 antibody also recognizes rhesus monkey and has inhibitor activity.
Applications Reported: This 14D3 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This 14D3 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated human peripheral blood cells using the Intracellular Fixation and Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Furthermore, due to the intracellular localization of a large portion of CTLA-4, for complete detection it may be necessary to assess intracellular expression, in addition to surface expression of CTLA-4.
eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome.
Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser.
Filtration: 0.2 µm post-manufacturing filtered.
CTLA4 (Cytotoxic T-Lymphocyte Antigen 4) is a CD28-family receptor expressed on mainly CD4+ T cells. It binds the same ligands as CD28 (CD80 and CD86 on B cells and dendritic cells), but with higher affinity than CD28. However, in contrast to CD28 which enhances cell function when bound at the same time as the T cell receptor, CTLA4 inhbits the T cell and prevents it from functioning. Intracellular CTLA4 is also found in regulatory T cells and may be important to their function.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: CD152; CD152 isoform; celiac disease 3; CTLA-4; cytotoxic T lymphocyte associated antigen 4 short spliced form; Cytotoxic T-lymphocyte protein 4; Cytotoxic T-lymphocyte-associated antigen 4; cytotoxic T-lymphocyte-associated protein 4; cytotoxic T-lymphocyte-associated serine esterase-4; insulin-dependent diabetes mellitus 12; ligand and transmembrane spliced cytotoxic T lymphocyte associated antigen 4
Gene Aliases: ALPS5; CD; CD152; CELIAC3; CTLA-4; CTLA4; GRD4; GSE; IDDM12
UniProt ID: (Human) P16410
Entrez Gene ID: (Human) 1493
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