Fc Receptor Blocker
Non-specific binding mediated by the Fc receptor is blocked by the inhibitor of binding to the Fc receptor. PHP-1 cells were either left untreated (purple histogram) or treated with Fc receptor binding inhibitory antibody (blue histogram) for 15 minutes at 4 ° C. The cells were then stained with 0.25 µg of Syrian hamster isotype control, APC (product # 17-4914-81). The grey histogram represents the autofluorescence of the unstained cells. Viable cells were used for analysis as determined by staining with fixable viability dye 780 (product # 65-0865).
- Flow cytometry (flow)
- Functional test (FN)
Proven dilution: 20 µL / test
Species reactivity: Human
Human, mouse, zebrafish
Host/isotype: This does not apply
Concentration: 1 mg / mL
Purification: Affinity chromatography
Storage buffer: PBS, pH 7.2
Contains: 0.09% sodium azide
Storage conditions: 4 ° C
Description: The human FcgammaR binding inhibitor can be used to inhibit the non-specific binding of the Fc-gamma receptor (FcgammaR) of mouse monoclonal antibodies used for flow cytometric analysis of human tissue. Four different classes of Fcgamma receptors have been identified: FcgammaRI (CD64), FcgammaRII (CD32), and FcgammaRIII (CD16), and are expressed at different levels in multiple cell lines, including high expression in myeloid, granulocytic, and B cells.
The biological function of FcgammaR, including initiation of endocytosis, phagocytosis, and antigen presentation, is triggered upon host immunoglobulin binding. The degree to which mouse monoclonal antibodies will bind to human FcgammaR varies depending on the isotype of the monoclonal antibody. Furthermore, different monoclonal antibodies of the same isotype will show different binding to human FcgammaRs.
Informed Applications: The purified Fc receptor binding inhibitor has been reported to be used in flow cytometric analysis. To inhibit non-specific binding of mouse monoclonal antibodies, add human Fc receptor binding inhibitor to samples and incubate on ice for 20 minutes. Without washing proceed to stain with primary antibody.
Tested Applications: The purified human Fc gamma binding inhibitor has been previously assessed and tested by inhibiting the binding of fluorochrome-conjugated isotype controls to U937 cells. This can be used at 20 µL per test. A test is defined as the amount (µg) of antibody that stains a cell sample in a final volume of 100 µL. The number of cells must be determined empirically, but can range from 10 ^ 5 to 10 ^ 8 cells/test.
Purity: greater than 90%, as determined by SDS-PAGE.
Aggregation: Less than 10%, as determined by HPLC.
Filtration: 0.2 µm post-fabrication filtering.
Four different classes of Fc receptors have been identified: FcyRI (CD64), FcyRII (CD32), and FcyRIII (CD16), and are expressed at varying levels in multiple cell lines, including high expression in myeloid, granulocytic, and B cells. Function Biological FcyR, which includes the initiation of endocytosis, phagocytosis, and antigen presentation, is triggered upon binding of host immunoglobulin. The degree to which mouse monoclonal antibodies will bind to human FcyR varies depending on the isotype of the monoclonal antibody. Furthermore, different monoclonal antibodies of the same isotype will show different binding to human FcyRs.