Description
The SE5A5 monoclonal antibody specifically binds to a common epitope on CD172a/SIRPα (90 kDa) and CD172b/SIRPβ1 (50 kDa). These transmembrane glycoproteins are members of the Signal Regulatory Protein (SIRP) family that, in turn, belongs to the Immunoglobulin superfamily. The SIRP family is comprised of two subgroups, SIRPα and SIRPβ that are distinguished by the presence (α) or absence (β) of a cytoplasmic domain containing two immunoreceptor tyrosine-based inhibition motifs (ITIM). CD172a/SIRPα is expressed on CD34+ stem/progenitor cells, cardiomyocytes, monocytes, macrophages, granulocytes, dendritic cells, and in the central nervous system. It binds to CD47 and is implicated in mediating inhibitory signals via the ITIM/SHP-2 association. CD172b/SIRPβ1 does not possess a cytoplasmic domain but instead the transmembrane domain contains a positively-charged residue that can interact with another transmembrane protein, DAP-12/KARAP. DAP-12 has two immunoreceptor tyrosine-based activation motifs (ITAM) within its cytoplasmic domain that are thought to link CD172b to cellular activation signaling. CD172b is expressed on myeloid cells, including peripheral blood monocytes and granulocytes. It is not expressed on CD34+ cells. CD172a and CD172b have complementary roles in signal regulation and may work together in tuning certain cellular responses to stimuli.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. An isotype control should be used at the same concentration as the antibody of interest.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
