Microplate assays to measure total and phospho-STAT6 in whole cells.
Thermo Scientific Pierce STAT6 In-Cell ELISA Kits are colorimetric and fluorescent microplate assays optimized to measure total and phosphorylated signal transducers and activators of transciption 6 (phospho-STAT6) in whole cells.
These ELISA kits for whole cells include a mouse monoclonal anti-STAT6 antibody, rabbit monoclonal anti-phospho-(Tyr641) STAT6 antibody, detection reagents and all buffers required for either colorimetric or near infrared detection. The STAT6 In-Cell ELISA Kits enable simultaneous detection of both total and phospho-(Tyr641) STAT6.
Measures total and phosphorylated signal transducers and activators of transciption 6 (STAT6) in whole cells using a 96- or 384-well microplates
colorimetric detection kit for use with standard microplate readers
near infrared fluorescent detection kit for use with IR imaging system
Whole-cell detection requires no cell lysis, protein extraction or special sample preparation
Provides better data and higher throughput than quantitative Western blotting
Less costly (one well vs. one lane) than Western blotting
Stimulation with EGF causes a 3-fold increase in the level of STAT6 phosphorylation A431 cells. Relative total and phospho-STAT6 levels were measured in A431 cells using the STAT6 Colorimetric In-Cell ELISA Kit. Cells were serum-starved overnight and stimulated with 100ng/ml EFG for five minutes immediately prior to fixation.The level of STAT6 phosphorylation increased approximately 3-fold in response to EGF stimulation where as total STAT6 was not significantly affected. Note: Absolute absorbance values cannot be directly compared between phospho and total STAT6 experiments because they use antibodies with different sensitivities.
Treatment with EGF increases phosphorylation of STAT6 in A431 cells. Relative total and STAT6 levels were measured in A431 cells using the (A) STAT6 Near Infrared In-Cell ELISA Kit. Cells were serum-starved overnight and then treated with EGF for five minutes immediately prior to fixation. The EGF-induced phosphorylation of STAT6 is reduced in a dose-dependent manner when cell receive a two hour pre-treatment with the minute EGFR inhibitor, PD168393 (B, STAT6 Colorimetric In-Cell ELISA Kit). Note: Absolute absorbance values cannot be directly compared between phospho and total STAT6 experiments because they use antibodies with different sensitivities.
Overview of STAT6 and phosphorylation of STAT6:
STAT6 belongs to the STAT family of transcription factors which regulate many aspects of cell growth, survival and differentiation. Inactive STAT proteins are located in the cytosol until phosphorylated by receptor associated kinases. Upon phosphorylation, STAT proteins form homodimers or heterodimers with other STAT proteins and translocate into the cell nucleus. STAT6 is known interact with the transcriptional regulators EP300, CREB, NFκB and is integral in the anti-apoptotic effects of IL-4 through regulation of BCL2L1/BCL-X(L) expression. The STAT6 In-Cell ELISA Kit enables simultaneous detection of both total and phospho-(Tyr641) STAT6.