Product Information

Contents: Functional Grade Purified anti-mouse CD152 (CTLA-4)
Catalog Number: 16-1521
Sizes: 50 ug, 100 ug, 500 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, No NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
KEEP CONTENTS STERILE.
Endotoxin Level: Less than 0.001 ng/ug antibody, as determined by the LAL assay.
Clone: 9H10
Isotype: Golden Syrian Hamster IgG

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-1521	Affinity Purified anti-mouse CD152 (CTLA-4, CTLA4)	N/A	N/A	FA FC IP
16-1521	Functional Grade* Purified anti-mouse CD152 (CTLA-4, CTLA4)	N/A	N/A	FA FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The 9H10 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 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.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The 9H10 antibody has been reported for use in flow cytometric analysis. It has also been reported for use in in vitro functional studies.

Applications Tested

The 9H10 antibody has been tested by flow cytometric analysis of resting and ConA-activated mouse splenocyte suspensions. This can be used at less than or equal to 0.5 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

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.

References

Krummel, M. F. and J. P. Allison (1995).CD28 and CTLA-4 have opposing effects on the response of T cells to stimulation. J Exp Med 182(2): 459-65.

Krummel, M. F. and J. P. Allison (1996). CTLA-4 engagement inhibits IL-2 accumulation and cell cycle progression upon activation of resting T cells. J Exp Med 183(6): 2533-40.
Chambers, C. A., M. F. Krummel, et al. (1996). The role of CTLA-4 in the regulation and initiation of T-cell responses.Immunol Rev 153: 27-46.

Lucy S. K. Walker, Helen E. Wiggett, Fabrina M. C. Gaspal, Chandra R. Raykundalia, Margaret D. Goodall, Kai-Michael Toellner, and Peter J. L. Lane. 2003. Established T Cell-Driven Germinal Center B Cell Proliferation Is Independent of CD28 Signaling but Is Tightly Regulated Through CTLA-4. J Immunol. 170:91-98.

Zheng SG, Wang JH, Stohl W, Kim KS, Gray JD, Horwitz DA. TGF-beta Requires CTLA-4 Early after T Cell Activation to Induce FoxP3 and Generate Adaptive CD4+CD25+ Regulatory Cells. J Immunol. 2006 Mar 15;176(6):3321-9. [9H10, in vivo neutralization, PubMed]

Sotomayor EM, Borrello I, et al. 1999. In vivo blockade of CTLA-4 enhances the priming of responsive T cells but fails to prevent the induction of tumor antigen-specific tolerance. Proc Natl Acad Sci U S A. 96(20):11476-81. (9H10, FA in vivo, PubMed)

Hurwitz AA, Yu TF, et al. 1998. CTLA-4 blockade synergizes with tumor-derived granulocyte-macrophage colony-stimulating factor for treatment of an experimental mammary carcinoma. Proc Natl Acad Sci U S A. 95(17):10067-71. (9H10, FA in vivo, PubMed)

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