• Assay Type

  Solid Phase Sandwich ELISA

• Format

  96-well strip plate

• Assay Length

  4 hours 40 mins (after plate preparation)

• Sample Type & Volume Required

  Cell lysates (100 μL)

• Range

  62.50 - 4,000 pg/mL

• Sufficient Materials

  Kits available for two, five, or fifteen 96-well plates*

• Specificity

  Please see the

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

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 # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na  ₂HPO  ₄, 1.5 mM KH  ₂O  ₄, pH 7.2 - 7.4, 0.2 μm filtered  
 
  Wash Buffer: (Catalog # ), or equivalent  
 
  Lysis Buffer*  
 
  IC Diluent*

Blocking Buffer*  
 
  Substrate Solution: 1:1 mixture of Color Reagent A (H  ₂O  ₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # )  
 
  Stop Solution: 2 N H  ₂SO  ₄ (Catalog # )  
 
  Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # ), or equivalent  
 
  Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent  
 

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

Preparation and Storage

• 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.

• Entrez Gene IDs:

  1027 (Human); 12576 (Mouse); 83571 (Rat);

• Long Name:

  p27 Cyclin Dependent Kinase 4 Inhibitor 1B

• Aliases:

  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