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Invitrogen
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Description
The monoclonal antibody B6H12 reacts to CD47 also known as integrin-associated protein (IAP), and neurophilin. This antibody has been reported to have neutralizing activity.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
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 (1.0 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.
Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.
Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.
Viability dye compatibility
• 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.
CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).
NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 636 nm; Emission: 727 nm; Laser: 633-640 nm (Red) Laser
CD47, known as integrin-associated protein (IAP), is a glycosylated transmembrane glycoprotein with five domains, expressed widely across hematopoietic cells like T and B cells, monocytes, platelets, and erythrocytes, as well as non-hematopoietic cells. It interacts with integrins such as CD51/CD61 and CD41/CD61, and serves as a receptor for thrombospondin, mediating bi-directional signaling that affects neural synaptic activity and macrophage phagocytosis. CD47 acts as a ligand for CD172a (SIRP alpha), an inhibitory receptor on macrophages, preventing phagocytosis of CD47-positive cells. This interaction influences cell migration, B cell adhesion, T cell activation, and neuronal development, particularly in synapse-rich brain and retina regions. It also modulates chondrocyte responses to mechanical signals. T cell expression of CD47 can lead to activation or apoptosis in the presence of thrombospondin. Monoclonal antibody stimulation of CD47 has been shown to induce CD4+CD25- suppressive activity and increase Foxp3 expression. CD47's role in membrane transport, signal transduction, and its broad tissue distribution highlight its significance in various physiological processes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
Protein Aliases: antigen identified by monoclonal antibody 1D8; Antigenic surface determinant protein OA3; CD47; CD47 antigen; CD47 antigen (Rh-related antigen, integrin-associated signal transducer); CD47 glycoprotein; IAP; integrin associated protein; Integrin-associated protein; integrin-associated signal transducer; Leukocyte surface antigen CD47; Protein MER6; Rh-related antigen; sCD47; soluble CD 47; soluble CD47
Gene Aliases: CD47; IAP; MER6; OA3
UniProt ID: (Human) Q08722
Entrez Gene ID: (Human) 961
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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