Description
The BNI3 monoclonal antibody specifically binds to the human cytolytic T lymphocyte-associated antigen (CTLA-4), also known as CD152. CTLA-4 is transiently expressed on activated CD28+ T cells and binds to CD80 and CD86 present on antigen presenting cells (APC) with high avidity. This interaction appears to deliver a negative regulatory signal to the T cell. Recent reports indicate that CTLA-4 is also expressed on B cells when cultured with activated T cells, suggesting a role for CTLA-4 in the regulation of B-cell response. Immobilized BNI3 antibody enhances T-cell proliferation induced by antibody-mediated crosslinking of CD3 and CD28. Recent studies have shown that CD152 can be expressed by regulatory T (Treg) cells. After cellular fixation and permeabilization, the BNI3 antibody can stain intracellular CD152 expressed in T cells including Treg cells. Clone BNI3 was studied in the VI Leukocyte Typing Workshop.
Immunohistochemistry: The BNI3.1 clone reactive against human CD152 is tested for immunohistochemical staining of acetone-fixed frozen sections. Tissue tested was human spleen and tonsil. The antibody stains activated T- and B-lymphocytes. The isotype control recommended for use with this antibody is purified mouse IgG2a (Cat. No. 550339). For optimal indirect immunohistochemical staining, the BNI3.1 antibody should be titrated (1:10 to 1:50 dilution) and visualized via a three-step staining procedure in combination with polyclonal, biotin conjugated anti-mouse Igs (multiple adsorbed) (Cat. No. 550337) as the secondary antibody and Streptavidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880).
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
3. An isotype control should be used at the same concentration as the antibody of interest.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
6. This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
8. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
10. For U.S. patents that may apply, see bd.com/patents.
