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 0.125 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. Filtration: 0.2 µm post-manufacturing filtered.
|OVA257-264 (SIINFEKL) peptide bound to H-2Kb|
|4° C, store in dark, DO NOT FREEZE!|
|Flow Cytometry, Immunohistochemistry|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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