Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

PathScan® Total p27 Kip1 Sandwich ELISA Kit #7768

When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

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.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Description

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.

ELISA

ELISA

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

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.

Background

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).

  1. Lloyd, R.V. et al. (1999) Am. J. Pathol. 154, 313-323.
  2. Polyak, K. et al. (1994) Genes Dev. 8, 9-22.
  3. Kato, J.Y. et al. (1994) Cell 79, 487-496.
  4. Vlach, J. et al. (1997) EMBO J. 16, 5334-5344.

Application References

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Protocols

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For Research Use Only. Not For Use In Diagnostic Procedures.

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