CD312 (EMR2) is a glycoprotein belonging to class B of the EGF-TM7 family of adhesion-type receptors. CD312 consists of a long extracellular N-terminal region with five EGF-like domains and a mucin stalk followed by seven transmembrane segments. EMR is cleaved at the juxtamembrane G-protein proteolytic site (GPS) and is expressed as a noncovalent heterodimer. CD312 is preferentially expressed on activated monocytes, macrophages and neutrophils. It localizes to the leading edge of migrating neutrophils and plays an important role in migration, adhesion and superoxide production. The portion of human CD312 N-terminal to the GPS domain (aa1-478) shares 64%, 59%, 48% and 45% aa identity with corresponding regions of canine CD312, equine CD312, mouse CD97 and rat CD97, respectively. Alternate splicing of human CD312 creates isoforms that contain 2-5 EGF-like domains
Suitable for use in Flow Cytometry. Other applications not tested.
Flow Cytometry: 10ul labeles 1-2.5x10e5 cells. Vaildated using whole blood monocytes.
Optimal dilutions to be determined by the researcher.
Flow Cytometry Protocol:
1. Cells may be Fc-blocked with 1ug of human IgG/10e5 cells for 15 minutes at RT. Do not wash excess blocking IgG from this reaction.
2. After blocking, add 10ul of C2549-36 to (same/less than)1x10e6 cells and incubate for 30 minutes at RT.
3. Remove unbound antibody by washing the cells twice in Flow Cytometry Staining Buffer (Catalog #F4568-10). Note that whole blood requires a RBC lysis step at this point using Flow Cytometry Human Lyse Buffer.
4. Resuspend the cells in Flow Cytometry Staining Buffer for analysis. As a control, cells in a separate tube should be treated with APC-labeled mouse IgG2b antibody.
Storage and Stability:
May be stored at 4 degrees C before opening. DO NOT FREEZE! Stable at 4 degrees C. Stable for at 12 months at 4 degrees C. Freezing allophycocyanin conjugates will result in a substantial loss of activity. Allophycocyaninconjugates are sensitive to light.