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Human Phospho-Histone H2AX (S139) DuoSet IC ELISA

R&D Systems, part of Bio-Techne | Catalog # DYC2288-2

R&D Systems, part of Bio-Techne
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DYC2288-2
DYC2288-5
DYC2288E

Ancillary Products Available

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Sample Type

Cell Lysates

Reactivity

Human

Human Phospho-Histone H2AX (S139) 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

Product Summary for Human Phospho-Histone H2AX (S139) DuoSet IC ELISA

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated human Histone H2AX (S139) in cell lysates. An immobilized capture antibody specific for Histone H2AX binds both phosphorylated and unphosphorylated Histone H2AX . After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard 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

Label

HRP

Scientific Data Images for Human Phospho-Histone H2AX (S139) DuoSet IC ELISA

Standard Curve

The Human Phospho-Axl DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis.

The Human Phospho-Axl DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis.

A172 human glioblastoma cells were treated with Gas-6-containing conditioned media1 from the human lung cell line, WI38, for fifteen minutes to induce tyrosine phosphorylation of Axl. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an Anti-Human Axl Monoclonal Antibody and Anti-Mouse IgG Agarose. Immunoblots were incubated with a Biotinylated Anti-Human Phospho-tyrosine Monoclonal Antibody (Catalog # BAM1676) to detect phospho-Axl. Bands were visualized with Streptavidin-HRP (Catalog # DY998) followed by chemiluminescent detection.
The Human Phospho-Axl DuoSet IC ELISA detects ligand-induced Axl tyrosine phosphorylation.

The Human Phospho-Axl DuoSet IC ELISA detects ligand-induced Axl tyrosine phosphorylation.

A172 cells were untreated or treated with Gas-6-containing conditioned media1 from the human lung cell line, WI38, for 15 minutes. ELISA and IP-Western blot (inset) analyses were done using 100 µg and 400 µg of lysate, respectively. IP-Western blots for phospho-Axl (p-Axl) were done as described in Figure 1. Blots were stripped and total Axl (Axl) was detected using a Biotinylated Anti-Human Axl Polyclonal Antibody (Catalog # BAF154).

Kit Contents for Human Phospho-Histone H2AX (S139) 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

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: Histone H2AX

Histone H2AX is one of a number of core histone proteins. In the cellular response to genotoxic insults, ATM and related protein kinases phosphorylate the carboxyl-terminal tail of the H2AX protein (gamma-H2AX). gamma-H2AX marks the site of damage and provides a nucleation site for the formation of damage response and repair complexes. BRCA1 carboxyl terminal (BRCT) domain-containing proteins are recruited to gamma-H2AX to create a protein scaffold for further assembly of signaling complexes.

Alternate Names

H2AFX

Entrez Gene IDs

3014 (Human)

Gene Symbol

H2AX

Additional Histone H2AX Products

Product Documents for Human Phospho-Histone H2AX (S139) DuoSet IC ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Phospho-Histone H2AX (S139) DuoSet IC ELISA

For research use only

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