Antibodies were obtained from the following suppliers: anti-human CD3 (BD Biosciences, 555335), anti-human CD8 (BD Biosciences 555366), anti-human CD107a (BD Biosciences 555801), anti-human CD137 (BD Biosciences 555956). Cell surface expression of ErbB2 was detected by biotylated anti-ErbB2 Affibody (Abcam, ab31890), and EGFR by FITC conjugated anti-EGFR affibody (Abcam, ab81872). EGFR, ErbB2 and CD19 specific CAR expression were detected by biotin-labeled polyclonal anti-human F(ab)2 antibody for (EGFR CAR) or anti-mouse F(ab)2 antibody (for ErbB2 and CD19 CARs)(Jackson Immunoresearch). Samples were then stained with PE-conjugated anti-human IgG Fc Ab (eBioscience, 12-4998-82) or phycoerythrin-labeled streptavidin (eBioscience, 17-4317-82). Flow cytometry acquisition was performed on either a BD FacsCalibur or Accuri C6 Cytometer (BD Biosciences). Analysis was performed using FlowJo software (Treestar).
Phycoerythrin labeled streptavidin
Manufactured by Thermo Fisher Scientific
Phycoerythrin-labeled streptavidin is a fluorescent conjugate used in various biomolecular detection and labeling applications. Streptavidin, a tetrameric protein, binds strongly to the small molecule biotin, enabling the detection and localization of biotinylated molecules. The phycoerythrin fluorescent dye is covalently attached to the streptavidin, providing a bright signal for visualization and quantification purposes.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd107a, by BD (2 mentions)
Anti human cd137, by BD (2 mentions)
Facscalibur, by BD (2 mentions)
Anti human cd3, by BD (2 mentions)
Accuri c6 cytometer, by BD (2 mentions)
Anti human cd8, by BD (2 mentions)
Anti mouse f ab 2 antibody, by Jackson ImmunoResearch (2 mentions)
Ab81872, by Abcam (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
3 protocols using phycoerythrin labeled streptavidin
1
Multiparametric Flow Cytometry Characterization
2
T Cell Surface Marker Detection
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Antibodies were obtained as follows: anti-human CD3 (BD Biosciences, 555335), anti-human CD8 (BD Biosciences, 555366), anti-human CD107a (BD Biosciences, 555801), and anti-human CD137 (BD Biosciences, 555956). The antibodies were incubated with T cells at 4°C for 25 min and washed twice (PBS with 2% FBS). Mesothelin CAR, ErbB2 CAR, CD19 CAR, and OKT3-28BB expression were detected by biotin-labeled polyclonal anti-mouse F(ab)2 antibody (Jackson Immunoresearch). Samples were then stained with phycoerythrin-labeled streptavidin (eBioscience, 17-4317-82). Flow cytometry acquisition was performed on either a BD FACSCalibur or Accuri C6 Cytometer (BD Biosciences). Analysis was performed using FlowJo software (Treestar).
3
PD-L1 Expression by Flow Cytometry
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Cells were stained with biotinylated monoclonal antibodies against human PD-L1 (Biolegend) and with phycoerythrin-labeled streptavidin (eBiosciences) for flow cytometric analysis with Cytoflex instrument (Beckman coulter). The data were analyzed using FlowJo software.
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