Anti-Paired Immunoglobulin-like Receptor B (PIR-B) (PE)

Mouse PIR-B is a 125kD type I transmembrane glycoprotein with six Ig-like domains in its extracellular domain (ECD) and four ITIM-like sequences in its cytoplasmic domain. The ECD of PIR-B is highly homologous to the ECDs of multiple mouse PIR-A receptors (92-99% aa sequence homology), which have short cytoplasmic tails lacking ITIM motifs. PIR-A receptors have a charged residue in their transmembrane domain that facilitates interaction with ITAM-containing adaptor molecules. Whereas PIR-A receptors deliver activation signals, PIR-B can inhibit receptor-mediated activation signaling. PIR-A and PIR-B have been shown to bind various mouse MHC class I molecules. They have been proposed to be orthologs of human leukocyte immunoglobulin- like receptors. Applications: Suitable for use in Flow Cytometry. Other applications not tested. Recommended Dilutions: Flow Cytometry: 10ul to label 1x10e6 mouse splenocytes, Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: This antibody has been tested for Flow Cytometry using mouse splenocytes using the following protocol. 1.Cells may be Fc-blocked with 1ug of mouse IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction. 2. After blocking, 10ul P2150-10B was added to up to 1-2.5x10e5 cells and incubated for 30 minutes at RT., 3. Unbound antibody was removed by washing the cells twice in F4568-10: Flow Cytometry Staining Buffer (1X). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer., 4. The cells were resuspended in F4568-10: Flow Cytometry Staining Buffer (1X) for final flow cytometric analysis. As a control for this analysis, cells in a separate tube should be treated with Rat IgG2a (PE) 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 12 months after receipt at 4°C. Freezing R-Phycoerythrin (PE) conjugates will result in a substantial loss of activity. PE conjugates are sensitive to light.