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 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. Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 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 (cat. SB-4400) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product No. 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product No. A10497). Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Somegeneralizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser HLA-DR, like other MHC class II molecules, is a transmembrane glycoprotein composed of a 36 kDa alpha chain (DRA) and 27 kDa beta chain (DRB). The alpha chain gene contains 5 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the two extracellular domains, and exon 4 encodes the transmembrane domain and the cytoplasmic tail. DRA does not have polymorphisms in the peptide binding part and acts as the sole alpha chain for DRB1, DRB3, DRB4 and DRB5. Within the DR molecule the beta chain contains all the polymorphisms specifying the peptide binding specificities. Hundreds of DRB1 alleles have been described and typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. HLA-DR is expressed primarily on antigen presenting cells such as B lymphocytes, monocytes, macrophages, thymic epithelial cells and activated T lymphocytes. Three loci, DR, DQ and DP, encode the major expressed products of the human class II region. The human MHC class II molecules bind intracellularly processed peptides, present them to T-helper cells, and have a critical role in the initiation of the immune response.
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 780|
|PBS with BSA and 0.09% sodium azide; pH 7.2|