BD, 555851, BD Pharmingen™ Purified Mouse Anti-Human CD152

Alternative Name: CTLA-4; AILIM; Cytotoxic T-lymphocyte protein 4
Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Isotype: Mouse BALB/c IgG2a, κ
Immunogen: Human CTLA4 Recombinant Protein
Application: Flow cytometry (Routinely Tested), Immunohistochemistry-frozen, Intracellular staining (flow cytometry) (Tested During Development)
Concentration: 0.5 mg/ml
Workshop Number: IX 34
Entrez Gene ID: 1493
RRID: AB_396174
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Recommended Assay Procedures: Recommended Assay Procedures For flow cytometric applications, a three step labeling procedure is recommended for amplifying signal. Suggested protocol for 3-step staining using concanavalin-A-stimulated peripheral blood mononuclear cells method: 1. Incubate 100 µl concanavalin-A-stimulated 3-day peripheral blood mononuclear cells (1 x 10^6) with primary (unconjugated) antibody for 20-30 minutes at room temperature. 2. Add 2 mls of 1X PharM Lyse (10X PharM Lyse, Cat. No. 555899) and incubate for 10-15 minutes. Centrifuge and aspirate. 3. Wash once with PBS/0.1% sodium azide/1% heat-inactivated  fetal bovine serum (PBS-FBS). Centrifuge and aspirate. 4. Add biotinylated goat anti-mouse Ig's  (Cat. No. 553999) and incubate for 20-30 minutes at room temperature. 5. Wash once with PBS-FBS. Centrifuge and aspirate. 6. Add SAV-PE (Cat. No. 554061) and incubate for 20-30 minutes in the dark at room temperature. 7. Wash once with PBS-FBS. Centrifuge and aspirate. Resuspend in 0.5 ml of PBS-FBS and analyze by flow cytometry.
Product Notices: Product Notices Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. Since applications vary, each investigator should titrate the reagent to obtain optimal results. An isotype control should be used at the same concentration as the antibody of interest. 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. 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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Please refer to http://regdocs.bd.com to access safety data sheets (SDS). For U.S. patents that may apply, see bd.com/patents.