Pan T cells were isolated from splenocytes using the Dynabeads FlowComp™ Mouse Pan T (CD90.2) kit. 200,000 mouse T cells and human PBMC, in complete RPMI were plated and rested for 2-3 hours at 37°C. Antibodies (mouse: αCD25PC61 or anti CD25NIB (both mIgG2a), αIL-2 neutralizing antibody (nAb) (JES6-1A12, BioXcell); human: RG6292, human IgG1 isotype control, Basiliximab (Absolute Antibody), Daclizumab (Absolute Antibody)) were added at 50 μg/ml (mouse) and 10 μg/ml (human), respectively, and were incubated with the cells for 30 mins at 37°C, following which cells were stimulated with IL-2 (50U/ml (mouse) and 10 U/ml (human) respectively, both Peprotech) for 10 mins at 37°C. IL-2 induced STAT5 phosphorylation was stopped when the cells were fixed and permeabilized with the eBioscience™ FoxP3 / Transcription Factor Staining Buffer Set and treated with the BD Phosflow Perm Buffer III. Blocking was calculated as follows: % blocking = 100 x [(% Stat5+ cells No Ab group - % Stat5+ cells 50ug/ml Ab group) / (% Stat5+ cells No Ab group)].
Nab jes6 1a12
Manufactured by BioXCell
The NAb) (JES6-1A12) is a lab equipment product used for the detection and quantification of specific proteins. It serves as a tool for researchers to study protein interactions and expression levels. The core function of this product is to provide a reliable and accurate method for analyzing target proteins in various biological samples.
Automatically generated - may contain errors
2 protocols using nab jes6 1a12
1
Quantifying IL-2 Signaling Blockade
2
Quantifying IL-2 Signaling Blockade
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Pan T cells were isolated from splenocytes using the Dynabeads FlowComp™ Mouse Pan T (CD90.2) kit. 200,000 mouse T cells and human PBMC, in complete RPMI were plated and rested for 2-3 hours at 37°C. Antibodies (mouse: αCD25PC61 or anti CD25NIB (both mIgG2a), αIL-2 neutralizing antibody (nAb) (JES6-1A12, BioXcell); human: RG6292, human IgG1 isotype control, Basiliximab (Absolute Antibody), Daclizumab (Absolute Antibody)) were added at 50 μg/ml (mouse) and 10 μg/ml (human), respectively, and were incubated with the cells for 30 mins at 37°C, following which cells were stimulated with IL-2 (50U/ml (mouse) and 10 U/ml (human) respectively, both Peprotech) for 10 mins at 37°C. IL-2 induced STAT5 phosphorylation was stopped when the cells were fixed and permeabilized with the eBioscience™ FoxP3 / Transcription Factor Staining Buffer Set and treated with the BD Phosflow Perm Buffer III. Blocking was calculated as follows: % blocking = 100 x [(% Stat5+ cells No Ab group - % Stat5+ cells 50ug/ml Ab group) / (% Stat5+ cells No Ab group)].
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