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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 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 & Permeabilization Buffer Set (Product # 88-8824-00 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 (0.03 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.
PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue 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.
仅用于科研。不用于诊断过程。未经明确授权不得转售。
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
蛋白别名: CD152; CD152 isoform; CD152 protein; celiac disease 3; CTLA-4; cytotoxic T lymphocyte associated antigen 4 short spliced form; Cytotoxic T-lymphocyte protein 4; cytotoxic T-lymphocyte protein 4 isoform CTLA4-TM; Cytotoxic T-lymphocyte-associated antigen 4; cytotoxic T-lymphocyte-associated protein 4; cytotoxic T-lymphocyte-associated serine esterase-4; ICOS; insulin-dependent diabetes mellitus 12; ligand and transmembrane spliced cytotoxic T lymphocyte associated antigen 4
基因别名: ALPS5; CD; CD152; CELIAC3; CTLA-4; CTLA4; EGK_04712; GRD4; GSE; IDDM12
UniProt ID: (Human) P16410
Entrez Gene ID: (Human) 1493, (Rhesus monkey) 705673