BD, 555850, BD Pharmingen™ Purified NA/LE 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), Functional assay (Tested During Development)
Concentration: 1.0 mg/ml
Workshop Number: IX 34
Entrez Gene ID: 1493
RRID: AB_396173
Storage Buffer: No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2 µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.
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. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
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 (1x10^6) with primary (unconjugated) antibody for 20-30 minutes at room temperature. 2. Add 2 mls of 1X lysing buffer (10X Pharm Lyse ™ Lysing Buffer, Cat. No. 555899) and incubate for 10-15 minutes. Centrifuge and aspirate. 3. Wash once with PBS/0.1% sodium azide/1% heat-activated fetal bovine serum (PBS-FBS). Centrifuge and aspirate. 4. Add biotinylated goat anti-mouse Ig  (Cat. No. 553999) and incubate for 20-30 minutes at room temperature. 5. Wash once with PBS-FBS. Centrifuge and aspirate. 6. Add PE-Streptavidin (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.