BD, 562260, BD Pharmingen™ PE 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_10895568
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Recommended Assay Procedures: Recommended Assay Procedures Suggested Staining Procedures for PE Mouse anti-Human LAP Antibody: 1.  Harvest the 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). 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 Mouse anti-Human CD4 (Cat. No. 347324) and either with the PE Mouse anti-Human LAP antibody (Cat. No. 562260) or with PE Mouse IgG1, κ Isotype control (Cat. No. 555749) for 30 minutes on ice, protected from light. 4.  Wash cells twice with stain buffer. 5.  Stain for Alexa Fluor® 488 Mouse anti-Human FoxP3, refer to Technical Data Sheet of Cat. No. 560047 for 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 anti-FoxP3 antibody for 30-45min. d.  Wash cells twice with stain buffer and acquire on the Flow cyotmeter.
Product Notices: Product Notices This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test). An isotype control should be used at the same concentration as the antibody of interest. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.