The monoclonal antibody B6H12 reacts to CD47 also known as integrin-associated protein (IAP), and neurophilin. CD47 is a glycosylated five transmembrane protein with a small alternatively spliced cytoplasmic domain. CD47 is involved in adhesion through interactions with SIRP (signal regulator protein) and is non-covalently associated with beta3 integrins CD51/CD61 and CD41/CD61. Furthermore this interaction can mediate bi-directional signaling to modify neural synaptic activity and regulate the phagocytic activities of macrophages. CD47 is the receptor for thrombospondin. T cell expression of CD47 can mediate activation or apoptosis (in the presence of high levels of thrombospondin). Recently stimulation of CD47 by monoclonal antibody has been shown to induce CD4+CD25- suppressive activity also increasing expression of Foxp3. Expression is found in the majority of hematopoietic cells including T and B cells, monocytes, platelets and erythrocytes (as part of the Rh complex). Expression is also found in non-hematopoietic cells.

This antibody has been reported to have neutralizing activity.

Applications Reported: This B6H12 antibody has been reported for use in flow cytometric analysis, immunoprecipitation, western blotting, and immunohistochemical staining of formalin-fixed paraffin embedded tissue sections.

Applications Tested: This B6H12 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (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.

NovaFluor dyes are not compatible with DNA intercalating viability dyes. Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100µL of cell sample containing 10^3 to 10^8 cells.

NovaFluor conjugates are based on Phiton™ technology utilizing novel nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 509 nm; Emission: 665 nm; Laser: 488 nm (Blue) Laser