Human Total p27/Kip1 DuoSet IC ELISA

Name: Human Total p27/Kip1 DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC2256-2

R&D Systems, Inc. a Bio-Techne Brand

Catalog # DYC2256-2

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Key Product Details

Assay Type:	Solid Phase Sandwich ELISA
Assay Range:	62.5 - 4,000 pg/mL
Sample Types:	Cell Lysates
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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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 for Human Total p27/Kip1 DuoSet IC ELISA

Assay Format	96-well strip plate (sold separately)
Sample Volume Required	Cell lysates (100 µL)
Reactivity	Human
Specificity	Please see the product datasheet
Detection Method	Colorimetric ELISA - 450nm (TMB)
Conjugate	Biotin
Label	HRP
Measurement	Quantitative ELISA
Sufficient Materials	Kits available for two, five, or fifteen 96-well plates*

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

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

Scientific Data Examples for Human Total p27/Kip1 DuoSet IC ELISA

	Standard Curve

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.

Item	Value
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 p27, KIP1P27KIP1, Kip1, MEN1B, MEN4, cyclin-dependent kinase inhibitor 1B, cyclin-dependent kinase inhibitor 1B (p27, Kip1), p27, p27Kip1

Citations for Human Total p27/Kip1 DuoSet IC ELISA (1)

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Citations are publications that use Bio-Techne products. Selected citations for Human Total p27/Kip1 DuoSet IC ELISA include:

Species: Human
Sample Types: Cell Lysates

Moss SC et al. (2010), Rapamycin regulates endothelial cell migration through regulation of the cyclin-dependent kinase inhibitor p27Kip1. J. Biol. Chem., 285(16):11991-7.
PMID: 20097763

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