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Pe conjugated anti rabbit igg h l secondary antibody

Manufactured by Cell Signaling Technology
Sourced in United States

The PE-conjugated anti-rabbit IgG (H+L) secondary antibody is a lab equipment product that is used to detect and visualize rabbit primary antibodies in various immunoassays. The antibody is conjugated with the fluorescent dye Phycoerythrin (PE), which allows for the detection of the bound rabbit primary antibodies.

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2 protocols using pe conjugated anti rabbit igg h l secondary antibody

1

Flow Cytometry Microglia Immunophenotyping

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Flow cytometry analysis of immunostained cells was performed following standard cell protocols. Prior to antibody labelling, the cell suspensions were incubated with anti-murine CD16/CD32 FC-Receptor (1:100, Cat. #14-0161-81, eBioscience, CA, USA) blocking reagent at 4 °C for 10 min. After blocking, the microglia were stained with FITC-conjugated mouse anti-rat CD11b (1:100, Cat. #554982, BD Biosciences, USA) and PerCP/Cy5.5-conjugated anti-rat CD45 (1:50, Cat. #202220, Biolegend, CA, USA). The microglia were then fixed and permeabilized with BD fixation/permeabilization solution (Cat. #554714, BD Cytofix/Cytoperm™, USA) for 20 min. The microglia were washed with BD Perm/Wash buffer (Cat. #554714, BD Cytofix/Cytoperm™, USA), resuspended in BD Perm/Wash buffer, and incubated with anti-iNOS (1:100, Cat. #ab15323, Abcam, UK) and rabbit mAb anti-arginase-1 (Arg-1) (1:100, Cat. # 3668s, Cell Signaling Technology, USA) primary antibodies for 30 min followed by a PE-conjugated anti-rabbit IgG (H+L) secondary antibody (1:100, Cat. #8885s, Cell Signaling Technology, USA). The cells were analyzed using a CytoFLEX instrument (Beckman Coulter Biotechnology, SuZhou). Ten thousand events were recorded, and microglia were identified by CD11b+/CD45low expression [33 (link)]. The results were analyzed using CytExpert software (Beckman Coulter Biotechnology, SuZhou).
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2

Microglial Immunophenotyping by Flow Cytometry

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Flow cytometry analysis of immunostained cells was performed following standard cell protocols. Prior to antibody labelling, the cell suspensions were incubated with anti-murine CD16/CD32 FC-Receptor blocking reagent at 4 °C for 10 min (eBioscience, CA, USA). After blocking, the microglia were stained with FITC-conjugated mouse anti-rat CD11b (BD Biosciences, USA) and PerCP/Cy5.5-conjugated anti-rat CD45 (Biolegend, CA, USA). The microglia were then xed and permeabilized with BD Fixation/Permeabilization buffer for 20 min. The microglia were washed with BD Perm/Wash buffer, resuspended in BD Perm/Wash buffer, and incubated with anti-iNOS (Abcam, UK) and rabbit mAb anti-arginase-1 (Arg-1) (Cell Signaling Technology, USA) primary antibodies for 30 min followed by a PE-conjugated anti-rabbit IgG (H + L) secondary antibody (Cell Signaling Technology, USA). The cells were analysed using a CytoFLEX instrument (Beckman Coulter Biotechnology, SuZhou). The results were analysed using CytExpert software (Beckman Coulter Biotechnology, SuZhou).
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