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Description: The UC10-4B9 monoclonal antibody reacts with mouse CD152, also known as the cytotoxic T lymphocyte antigen-4 (CTLA-4). CTLA-4, a protein with structural similarities to CD28, is expressed on activated T cells at low level 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 a large proportion of newly synthesized CTLA-4 in response to TCR signaling to be localized intracellularly.
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.
Applications Reported: This UC10-4B9 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This UC10-4B9 antibody has been tested by intracellular staining followed by flow cytometric analysis of mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins" located at Flow Protocols. This may be used at less than or equal to 1.0 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Brilliant Ultra Violet™ 737 (BUV737) is a tandem dye that emits at 732 nm and is intended for use on cytometers equipped with an ultraviolet (355 nm) laser. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright, Brilliant Violet, Brilliant Ultra Violet, or other polymer dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401-42) or Brilliant Stain Buffer™ (Product # 00-4409-75) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer or Brilliant Stain Buffer for more information.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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.
Our internal testing suggests that Brilliant Ultra Violet™ 737 (BUV737) is compatible with short-term methanol-based fixation, but should not be stored in buffers containing methanol for longer than one hour.
Excitation: 355 nm; Emission: 732 nm; Laser: Ultraviolet Laser
BRILLIANT ULTRA VIOLET™ is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by Sirigen.™
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.
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.
Protein Aliases: CD152; CD152 antigen; CTLA-4; Cytotoxic T-lymphocyte protein 4; Cytotoxic T-lymphocyte-associated antigen 4; ICOS
Gene Aliases: Cd152; Ctla-4; Ctla4; Ly-56
UniProt ID: (Mouse) P09793
Entrez Gene ID: (Mouse) 12477
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