Anti-Latency Associated Peptide, Transforming Growth Factor beta1 (LAP, TGFb1) (PE)

TGF-beta is secreted by cells in the form of an inactive complex. This complex consists of TGF-beta associated non-covalently with a protein designated the latency associated peptide (LAP). TGF-beta and LAP represent components of a pro-peptide that is cleaved in a post-golgi compartment prior to secretion. LAP and TGF-beta each consist of a disulfide-linked homodimer and the association of these two components renders TGF-beta inactive and inaccessible to anti-TGF-beta antibodies. Applications: Suitable for use in Flow Cytometry. Other applications not tested. Recommended Dilution: Flow Cytometry: 10ul labels <1x10e6 cells. Validated using CD4+CD25+ human PBMCs and mouse splenocytes., Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Cells may be Fc-blocked with 1ug of human or mouse IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction., 2. After blocking add 10ul of L1393-30 to <1x10e6 cells and incubate for 30 minutes at RT., 3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (Catalog # F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human or Mouse Lyse Buffer., 4. Resuspend the cells in Flow Cytometry Staining Buffer for final flow cytometric analysis. As a control for analysis, cells in a separate tube should be treated with PE-labeled mouse IgG1 antibody. Storage and Stability: May be stored at 4°C before opening. DO NOT FREEZE! Stable at 4°C as an undiluted liquid. Stable for at least 12 months at 4°C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.