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.
Human Total p27/Kip1 DuoSet IC ELISA
R&D Systems | Catalog # DYC2256-2
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
62.5-4000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human
Human Total p27/Kip1 DuoSet IC ELISA 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
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Product Summary for Human Total p27/Kip1 DuoSet IC ELISA
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 Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human Total p27/Kip1 DuoSet IC ELISA
Standard Curve
Kit Contents for Human Total p27/Kip1 DuoSet IC ELISA
- 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
Background: p27/Kip1
Long Name
p27 Cyclin Dependent Kinase 4 Inhibitor 1B
Alternate Names
CDKN1B, Kip1
Gene Symbol
CDKN1B
Additional p27/Kip1 Products
Product Documents for Human Total p27/Kip1 DuoSet IC ELISA
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Total p27/Kip1 DuoSet IC ELISA
For research use only
Related Research Areas
Citations for Human Total p27/Kip1 DuoSet IC ELISA
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
