Human Total p27/Kip1 DuoSet IC ELISA

Catalog # Availability Size / Price Qty
DYC2256E
DYC2256-5
DYC2256-2
Ancillary Products Available
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Human Total p27/Kip1 DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 mins (after plate preparation)
Sample Volume Required
Cell lysates (100 µL)
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total p27 / Kip1 in cell lysates. An immobilized capture antibody specific for p27 / Kip1 binds both phosphorylated and unphosphorylated p27 / Kip1. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15- (96-well) plate pack sizes
  • Economical alternative to Western blot

Kit Content

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control
  • Streptavidin-HRP

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*


Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Scientific Data

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Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: p27/Kip1

The cyclin-dependent kinase (cdk) inhibitor p27/Kip1 is a tumor suppressor protein that acts in the nucleus to enforce cell cycle checkpoints. p27/Kip1 binds to and inhibits many cyclin/cdk complexes, including cyclin D/cdk4, cyclin E/cdk2, and cyclin A/cdk2. This inhibition can subsequently block progression through different phases of the cell cycle. p27/Kip1 is phosphorylated at T198 in cells treated with the AMP-dependent protein kinase (AMPK) activator AICAR (5-amino-4-imidazolecarboxamide riboside) or glucose withdrawal.

Long Name:
p27 Cyclin Dependent Kinase 4 Inhibitor 1B
Entrez Gene IDs:
1027 (Human); 12576 (Mouse); 83571 (Rat)
Alternate Names:
CDKN1B; CDKN4; cyclin-dependent kinase inhibitor 1B (p27, Kip1); cyclin-dependent kinase inhibitor 1B; Cyclin-dependent kinase inhibitor p27; Kip1; KIP1P27KIP1; MEN1B; MEN4; p27; p27Kip1

Citation for Human Total p27/Kip1 DuoSet IC ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Rapamycin regulates endothelial cell migration through regulation of the cyclin-dependent kinase inhibitor p27Kip1.
    Authors: Moss SC, Lightell DJ, Marx SO, Marks AR, Woods TC
    J. Biol. Chem., 2010-01-22;285(16):11991-7.
    Species: Human
    Sample Types: Cell Lysates

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