Analysis Note

ControlMouse spleen cell lysate

Application

Anti-CTLA4 (CD152) Antibody, clone 9H10 is an antibody against CTLA4 (CD152) for use in WB, FC & IP.

Research Sub CategoryTranscription FactorsInflammation & Autoimmune Mechanisms

Flow Cytometry Analysis: Representative images from a previous lot of primary antibody that was biotinylated. Results with 04-963 are expected to be similar if used directly with an anti-Syrian hamster conjugated antibody, however because of the low expression of CTLA4 Fab fragmented goat anti-Syrian hamster biotin followed SA-fluorophore is recommended for best results.
FACS analysis of ConA (24 hours) stimulated BALB/C splenocytes for surface expression of CTLA-4 (CD152). Cells were stained with 10 ug of biotin-labeled hamster anti-CTLA-4 ,clone 9H10 (open histogram) or with PE-Sav secondary antibody alone (histogram, Figure B). Note the distinct signal difference between the control (dark control histogram) and test (clear histogram).
Immunoprecipitation: Clone 9H10 is widely used for immunoprecipitaiton of CTLA-4; recommended lysis buffers include 1% NP40; 0.5% triton X-100 with proteinase inhibitors; 1-5 µg of 9H10 is added to 300-500 µL of whole cell lysate or cell membrane protein preparations (250-500 mg/mL totall protein) and incubated 3-6 hours at 4°C, followed by capture with protein G beads following standard immunoprecipitation methods (Zhang, Y., et al., 1997).

Research CategoryInflammation & Immunology

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

CTLA4, Cytotoxic T lymphocyte antigen 4, also known as CD152, belongs to the family of immunoglobins. It is widely expressed on activated T lymphocytes and is involved in the maintenance of immune tolerance. CTLA4 is a homolog for CD28. They are able to bind to B7-1 and B7-2; however, CTLA4 is able to bind to both with a higher affinity. Defects in CTLA4 have been linked to Rheumatoid arthritis, Addisons disease, Graves disease and many other autoimmune diseases.

Immunogen

Mouse CTLA-4/human IgG1 fusion protein corresponding to residues surrounding CTLA4.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Purified

Purified hamster monoclonal IgG in buffer containing 0.1 M Tris-Glycine (pH 7.4) with 150 mM NaCl and 0.05% sodium azide.

Format: Purified

Quality

Evaluated by Western Blot in mouse spleen lysate.
Western Blot Analysis: 1 µg/mL of a biotinylated version of this antibody detected CTLA4 in 10 µg of mouse spleen lysate.
THIS ANTIBODY IS NO BIOTINYLATED.

Specificity

This antibody recognizes CTLA4. Antibody 9H10 is specific for CTLA-4 by four criteria: (a) Reactivity against CTLA-4 Ig but not CD4 lg; (b) the ability to block CTLA-4 Ig binding to B7 transfectants; (c) the ability to stain activated T cells but not freshly isolated T cells; and (d) the ability to stain a CTLA-4 transfectant but not control transfectants (Krummel, M.F., et al., 1995).

Storage and Stability

Stable at 2-8°C for 1 year from date of receipt.

Target description

~30 kDa