OVA257-264 (SIINFEKL) peptide bound to H-2Kb Monoclonal Antibody (eBio25-D1.16 (25-D1.16)), eBioscience™, Invitrogen™

Description

Description: The 25-D1.16 monoclonal antibody reacts with the ovalbumin-derived peptide SIINFEKL bound to H-2Kb of MHC class I, but not with unbound H-2Kb, or H-2Kb bound with an irrelevant peptide. This antibody has proven to be very useful tracking the quantity and localization of these specific antigen-presenting cells (APC) in vivo. Applications Reported: This eBio25-D1.16 (25-D1.16) antibody has been reported for use in flow cytometric analysis, and immunohistochemical staining. Applications Tested: This eBio25-D1.16 (25-D1.16) antibody has been tested by flow cytometric analysis of SIINFEKL-pulsed C57Bl/6 splenocytes. This can be used at less than or equal to 0.25 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Cells can be pulsed with the SIINFEKL peptide according to the following protocol: 1. With cells in flow staining buffer, add SIINFEKL peptide to a final concentration of 30 uM. 2. Incubate cells at 37°C for 2 hours. 3. Wash cells with flow staining buffer. 4. Proceed with cell surface staining as normal. For additional information see the references listed below. Purity: Greater than 90%, as determined by SDS-PAGE.