3.25 x 107 Dynabeads (Life Technologies) were separately coated with 6 ug/mL of the following antibodies: (a) anti-eNOS (NOS3 A9; Santa Cruz Biotech); (b) eNOS isotype control (IgG2a Clone DAK-GO5, Dako); (c) anti-PlAP antibody (NDOG2); and (d) PlAP isotype control (IgG1 Clone MOPC-21, BioLegend). STBMV or STBEX pooled from 4 NP (1 mg/mL) were incubated with anti-human Fc receptor blocking reagent (10 µL, Miltenyi Biotec) for 10 min at 4°C to block any non-specific antigen binding (same pool was used at each experiment). Next, antibody-coated Dynabeads were incubated overnight at 4°C with 25 µg of protein from either STBMV (~2.14 x 109 EV/mL) or STBEX (~3.2 x 109 EV/mL) pools in 1 mL filtered PBS, as per manufacturer’s instructions. STBMV or STBEX bound to antibody-coated Dynabeads were separated and washed with PBS using a magnetic particle concentrator (Dynal MPC-S, ThermoFisher) and processed for Western blotting. See Methods in the Online Data Supplement for percentage calculations.
Anti human fc receptor blocking reagent
Manufactured by Miltenyi Biotec
The Anti-human Fc receptor blocking reagent is a laboratory tool designed to block Fc receptor interactions. It is used to prevent non-specific binding of antibodies to Fc receptors, which can interfere with experimental results.
Automatically generated - may contain errors
2 protocols using anti human fc receptor blocking reagent
1
Antibody-based Extracellular Vesicle Isolation
2
Extracellular Vesicle Proteomic Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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3.25×107 Dynabeads (Life Technologies) were separately coated with 6 μg/mL of the following antibodies: (1) anti-eNOS (NOS3 A9; Santa Cruz Biotech); (2) eNOS isotype control (IgG2a Clone DAK-GO5; Dako); (3) anti-PlAP antibody (NDOG2); and (4) PlAP isotype control (IgG1 Clone MOPC-21; BioLegend). STBMV or STBEX pooled from 4 NP (1 mg/mL) were incubated with anti-human Fc receptor blocking reagent (10 μL; Miltenyi Biotec) for 10 minutes at 4°C to block any nonspecific antigen binding (same pool was used at each experiment). Next, antibody-coated Dynabeads were incubated overnight at 4°C with 25 μg of protein from either STBMV (≈2.14×109 EV/mL) or STBEX (≈3.2×109 EV/mL) pools in 1 mL filtered PBS, as per manufacturer’s instructions. STBMV or STBEX bound to antibody-coated Dynabeads were separated and washed with PBS using a magnetic particle concentrator (Dynal MPC-S; ThermoFisher) and processed for Western blotting. See Methods section is available in the online-only Data Supplement for percentage calculations.
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