Product Pathways - Cell Cycle / Checkpoint
PathScan® Total p27 Kip1 Sandwich ELISA Kit #7768
|7768S||1 Kit (96 assays)||---||In Stock||---|
|7768||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at email@example.com.
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|Kit Includes||Volume||Solution Color|
|p27 Kip1 Rabbit Ab Coated Microwells||96 tests|
|p27 Kip1 Mouse Detection Ab||11 ml||Green|
|Anti-Mouse IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
|ELISA Sample Diluent||25 ml||Blue|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
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 Cell Lysis Buffer, which is stored at –20°C (packaged separately).
The PathScan® Total p27 Kip1 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p27 Kip1. A p27 Kip1 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, p27 Kip1 protein is captured by the coated antibody. Following extensive washing, a p27 Kip1 Mouse Detection Antibody is added to detect the captured p27 Kip1 protein. Anti-Mouse 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 the developed color is proportional to the quantity of p27 Kip1 protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Total p27 Kip1 Sandwich ELISA Kit detects endogenous levels of total p27 Kip1 protein. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Figure 2. The relationship between the protein concentration of lysates from untreated and hTGF-β1-treated NIH/3T3 cells and the absorbance at 450 nm using the PathScan® Total p27 Kip1 Sandwich ELISA Kit is shown.
ELISA - Western correlation
Figure 1. The expression of p27 Kip1 in NIH/3T3 cells can be reduced when cells are treated with Human Transforming Growth Factor β1 (hTGF-β1) #8915, as detected by the PathScan® Total p27 Kip1 Sandwich ELISA Kit. NIH/3T3 cells (80% confluent) were treated with hTGF-β1 (2.5 μg/ml, 24 hr). The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blot using p27 Kip1 (SX53G8.5) Mouse mAb #3698 is shown in the bottom figure.
p27 Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. Like its relatives, p57 Kip2 and p21 Waf1/Cip1, the ability to enforce the G1 restriction point is derived from its inhibitory binding to CDK2/cyclin E and other CDK/cyclin complexes. Expression levels of p27 are upregulated in quiescent cells and in cells treated with cAMP or other negative cell cycle regulators. Downregulation of p27 can be induced by treatment with interleukin-2 or other mitogens; this involves phosphorylation of p27 and its degradation by the ubiquitin-proteasome pathway (1-4).
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PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.