Product Pathways - Jak/Stat Pathway
PathScan® Phospho-Stat6 (Tyr641) Sandwich ELISA Kit #7275
|7275C||1 Kit (96 assays)||---||In Stock||---|
|7275||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
|Kit Includes||Volume||Solution Color|
|Stat6 Mouse mAb coated microwells||96 tests|
|Phospho-Stat6(Tyr641) Rabbit Detection Antibody||1 Ea||Green (Lyophilized)|
|Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated)||1 Ea||Red (Lyophilized)|
|Detection Antibody Diluent||11 ml||Green|
|HRP Diluent||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Lysis Buffer, which is stored at –20°C (packaged separately).
The PathScan® Phospho-Stat6 (Tyr641) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat6 protein phosphorylated at Tyr641. A Stat6 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, Stat6 (phospho or nonphospho) protein is captured by the coated antibody. Following extensive washing, Phospho-Stat6 (Tyr641) Rabbit Detection Antibody is added to detect the captured phospho-Stat6 protein. Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of Stat6 phosphorylated at Tyr641.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Phospho-Stat6 (Tyr641) Sandwich ELISA Kit detects endogenous levels of phospho-Stat6 (Tyr641) in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1. Treatment of ACHN cells with Human Interleukin-4 (hIL-4) #8919 stimulates phosphorylation of Stat6 at Tyr641, as detected by PathScan® Phospho-Stat6 (Tyr641) Sandwich ELISA Kit #7275, but does not affect levels of total Stat6 protein detected by PathScan® Total Stat6 Sandwich ELISA Kit #7267. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using Stat6 Antibody #9362 (left panel) or Phospho-Stat6 (Tyr641) (C11A12) Rabbit mAb #9364 (right panel) are shown in the bottom figure.
Figure 2. Relationship between protein concentration of lysates from ACHN cells, untreated or treated with Human Interleukin-4 (hIL-4) #8919, and the absorbance at 450 nm is shown. Cells (80% confluence) were treated with hIL-4 (100 ng/ml) and lysed after incubation at 37ºC for 15-20 minutes.
Upon activation by Janus kinases, Stat6 translocates to the nucleus where it regulates cytokine-induced gene expression. Stat6 is activated via phosphorylation at Tyr641 and is required for responsiveness to IL-4 and IL-13 (1-4). In addition, Stat6 is activated by IFN-α in B cells, where it forms transcriptionally active complexes with Stat2 and p48 (5,6). Protein phosphatase 2A is also involved in regulation of IL-4-mediated Stat6 signaling (7).
- Nelms, K. et al. (1999) Ann. Rev. Immunol. 17, 701-738.
- Malabarba, M.G. et al. (1996) Biochem. J. 319, 865-872.
- Hou, J. et al. (1994) Science 265, 1701-1706.
- Quelle, F.W. et al. (1995) Mol. Cell. Biol. 15, 3336-3343.
- Takeda, K. et al. (1996) Nature 380, 627-630.
- Gupta, S. et al. (1999) J. Immunol. 163, 3834-3841.
- Woetmann, A. et al. (2003) J. Biol. Chem. 278, 2787-2791.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9998 BSA
- 9803 Cell Lysis Buffer (10X)
- 8919 Human Interleukin-4 (hIL-4)
- 7267 PathScan® Total Stat6 Sandwich ELISA Kit
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween® 20 (PBST-20X)
- 9364 Phospho-Stat6 (Tyr641) (C11A12) Rabbit mAb
- 9361 Phospho-Stat6 (Tyr641) Antibody
- 7002 STOP Solution
- 5397 Stat6 (D3H4) Rabbit mAb
- 9362 Stat6 Antibody
- 7004 TMB Substrate
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
PathScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.