BD, 562490, BD Horizon™ PE-CF594 Mouse Anti-Human LAP

Alternative Name: Latency-associated peptide; TGFB1; TGFbeta; Transforming growth factor beta
Reactivity: Human (QC Testing)
Isotype: Mouse BALB/c IgG1, κ
Immunogen: Human TGF-β1
Application: Intracellular staining (flow cytometry) (Routinely Tested), Flow cytometry (Tested During Development)
Vol. Per Test: 5 µl
RRID: AB_2737626
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.
Recommended Assay Procedures: Recommended Assay Procedures Suggested Staining Procedures for BD Horizon™ PE-CF594 Mouse Anti-Human LAP antibody: 1. Harvest PBMCs after stimulation (24 hours) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725) antibodies. 2. Wash the cells twice with stain buffer [eg. BD Pharmingen™ Stain Buffer (FBS), Cat. No. 554656]. 3. Stain 1 × 10^6 cells with PerCP-Cy™ 5.5 Mouse Anti-Human CD4 (Cat. No. 341654) and either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292) or BD Horizon™ PE-CF594 Mouse Anti-Human LAP antibody (Cat. No. 562490) for 30 minutes on ice, protected from light. 4. Wash cells twice with stain buffer. 5. Stain cells for intracellular FoxP3 with Alexa Fluor® 647 Mouse Anti-Human FoxP3 antibody; refer to the Technical Data Sheets of Cat. No. 560889 or 560045 for a detailed protocol. In brief, a. Add 2 ml of 1 × FoxP3 buffer A to the cell pellet. b. Centrifuge and incubate in 0.5 ml of buffer C for 30 minutes. c. Wash cells twice with stain buffer and stain with fluorescent Anti-FoxP3 antibody for 30-45min. d. Wash cells twice with stain buffer and analyze by flow cytometry.