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The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 421™ under optimal conditions. The solution is free of unconjugated Brilliant Violet 421™ and unconjugated antibody.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤5 µl per million cells or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.
The UC10-4B9 antibody can enhance T cell co-stimulation by blocking CTLA-4 interactions with the B7 co-receptors, favoring CD28 interactions. Additional reported applications (for the relevant formats) include: immunoprecipitation1, in vitro stimulation, in vitro and in vivo blocking1-4 of ligand binding, and as ELISA capture antibody5. To reduce non-specific binding to cells bearing Fc-receptors, pre-incubation of cells with anti-mouse CD16/CD32, clone 93 (Cat. No. 101301/101302), is recommended prior to immunofluorescent staining. For most successful immunofluorescent staining results, it may be important to maximize signal over background by using a relatively bright fluorochrome-antibody conjugate (Cat. No. 106306) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated anti-Armenian hamster IgG (Cat. No. 405501) second step, followed by SAv-PE (Cat. No. 405204)). The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 106308).
Application References:
1. Walunas TL, et al. 1994. Immunity 1:405. (Block IP) 2. Cilio CM, et al. 1998. J. Exp. Med. 188:1239. (Block) 3. Issazadeh S, et al. 1999. J. Immunol. 162:761. (Block) 4. McCoy K, et al. 1997. J. Exp. Med. 186:183. (Block) 5. Hsu HC, et al. 2007. J. Immunol. 178:5357. (ELISA Capture) 6. Sugita S, et al. 2010. Invest. Ophthalmol. Vis. Sci. 51:5783. PubMed
ConA stimulated C57BL/6 splenocytes (3 days) were stained with CD3 APC and CD152 (clone UC10-4B9) Brilliant Violet 421™ (top) or Armenian Hamster IgG Brilliant Violet 421™ isotype control (bottom).
CD152, also known as CTLA-4 or Ly-56, is a 33 kD member of the immunoglobulin superfamily. It is expressed on activated T and B lymphocytes. CD152 is similar to CD28 in amino acid sequence, structure, and genomic organization and these two receptors share common B7 family counter-receptors (B7-1, B7-2). Whereas CD28 delivers a costimulatory signal in T cell activation, CTLA-4 negatively regulates cell-mediated immune responses. CD152 is thought to play a role in the induction and maintenance of immunological tolerance as well as the development of protective immunity and thymocyte regulation.
Other Names:
Cytotoxic T Lymphocyte-Associated Antigen-4 (CTLA-4), Ly-56
Structure:
Ig superfamily, 33 kD
Distribution:
Activated T and B cells
Function:
Negative regulator of T cell activation
Ligand Receptor:
CD80 (B7-1), CD86 (B7-2)
Antigen References:
1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press. 2. Allison JP, et al. 1995. Science 270:932. 3. Waterhouse P, et al. 1995. Science 270:985. 4. Linsley PS, et al. 1991. J. Exp. Med. 174:561.
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This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
Biotin anti-mouse CD152
Con A-stimulated (day-2) Balb/c mouse splenocytes stained with biotinylated UC10-4B9, followed by Sav-PE
PE anti-mouse CD152
Con A-stimulated BALB/c splenocytes (day-2) were stained with CD3 APC and UC10-4B9 PE
APC anti-mouse CD152
ConA stimulated C57BL/6 splenocytes (3 days) stained with UC10-4B9 APC and CD3e (145-2C11) PE
ConA stimulated C57BL/6 splenocytes (3 days) stained with Armenian Hamster IgG isotype APC and CD3e (145-2C11) PE
Brilliant Violet 421™ anti-mouse CD152
ConA stimulated C57BL/6 splenocytes (3 days) were stained with CD3 APC and CD152 (clone UC10-4B9) Brilliant Violet 421™ (top) or Armenian Hamster IgG Brilliant Violet 421™ isotype control (bottom).
