Description: The J43 monoclonal antibody reacts with mouse PD-1 (programmed death-1), a 55 kDa member of the Ig superfamily. PD-1 contains the immunoreceptor tyrosine-based inhibitory motif (ITIM) and plays a key role in peripheral tolerance and autoimmune disease in mice. PD-1 is expressed mainly on activated T and B lymphocytes. Two novel B7 Family members have been identified as PD-1 ligands, PD-L1 (B7-H1) and PD-L2 (B7-DC). Evidence reported to date suggests overlapping functions for these ligands and their constitutive expression on some normal tissues and upregulation on activated antigen-presenting cells. It is reported that J43 inhibits the binding of mouse PD-L1-Ig and mouse PD-L2-Ig to PD-1/BHK transfected cells. When administrated in vivo, both intact and Fab of J43 are reported to enhance contact hypersensitivity and exacerbate acute GVHD similar to transfer of PD-1-deficient cells. Injection of J43 also exacerbates EAE and NOD diabetes as do specific antibodies to mouse PD-L1 and PD-L2. Applications Reported: This J43 antibody has been reported for use in flow cytometric analysis. Applications Tested: This J43 antibody has been tested by flow cytometric analysis of stimulated mouse splenocytes. This can be used at less than or equal to 1 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. Super Bright 702 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 702 nm. We recommend using a 710/50 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Staining Buffer (Product No. SB-4400) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 702 nm; Laser: Violet Laser HLA-class I antigens are widely distributed on human nucleated cells. The intensity of HLA-ABC maybe altered in pathological states e.g. maligt cells may lose HLA-ABC, whereas hepatocytes in alcoholic hepatitis, biliary cirrhosis and chronic active hepatitis may show enhanced expression of HLA-ABC.
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 702|
|PBS with BSA and 0.09% sodium azide; pH 7.2|