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EpiQuik In Situ DNA Damage Assay Kit

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Schematic procedure for using the EpiQuik In Situ DNA Damage Assay Kit.
MCF-7 cells in microplate wells were exposed to etoposide for 4 h and DNA damage was detected by the measuring level of phosphor-H2AX.
Input Type: Cells
Research Area: DNA Damage & Repair
Target Application: Amount Quantitation
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-6001-09696 assays $433.00 
P-6001-1922x96 assays $656.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

The EpiQuik™ In Situ DNA Damage Assay Kit is a convenient set of tools that allows the experimenter to detect DNA damage or apoptosis by measuring phosphorylation of H2AXSer139 in situ. The kit is ready-to-use and provides all the essential components needed for specifically measuring DNA damage in situ through phospho H2AXSer139 detection using cultured adherent cells. The kit has the following advantages:

  • Quick and efficient procedure, which can be finished within 3 hours.
  • Innovative colorimetric assay without the need for radioactivity, electrophoresis, and chromatography.
  • Measurement of in situ histone H2AX phosphorylation without the need to prepare cell lysates.
  • Microplate format makes the assay suitable for high throughput analysis of agents that increases or inhibits DNA damage.
  • Simple, reliable, and consistent assay conditions.

Principle & Procedure
EpiQuik™ In Situ DNA Damage Assay Kit is a whole cell-based detection of DNA damage and/or apoptosis. In this assay, adherent cells are cultured in conventional 96 -well microplates. After your experimental treatment, cells are fixed and permeabilized. The phosphorylation of H2AX at serine139, the sensitive marker of DNA damage, is then detected by an anti-phospho H2AXSer139 antibody. The ratio or amount of phospho H2AXSer139 can be quantified through HRP conjugated secondary antibody-color development system and is proportional to the intensity of color development.

 
Fig. 1. Schematic procedure for using the EpiQuik In Situ DNA Damage Assay Kit.

Fig. 2. MCF-7 cells in microplate wells were exposed to etoposide for 4 h and DNA damage was detected by the measuring level of phosphor-H2AX.

Product Components

A1 (10X Wash Buffer)
A2 (Permeabilizing Buffer)
A3 (Blocking Buffer)
A4 (Antibody Buffer)
A5 (Capture Antibody, 100 µg/ml)*
A6 (Detection Antibody, 200 µg/ml)*
A7 (Developing Solution)
A8 (Stop Solution)
30% H2O2 Solution
Positive Control (Etoposide, 10 mM)*
Microplates
User Guide

* For maximum recovery of the products, centrifuge the original vial prior to opening the cap.

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

Festuccia C et. al. (February 2018). The first-in-class alkylating deacetylase inhibitor molecule tinostamustine shows antitumor effects and is synergistic with radiotherapy in preclinical models of glioblastoma. J Hematol Oncol. 11(1):32.

RuiXie et. al. (August 2017). Rational design, synthesis and preliminary antitumor activity evaluation of a chlorambucil derivative with potent DNA/HDAC dual-targeting inhibitory activity Bioorganic & Medicinal Chemistry Letters. 27(16)

Gravina GL et. al. (June 2017). The novel CXCR4 antagonist, PRX177561, reduces tumor cell proliferation and accelerates cancer stem cell differentiation in glioblastoma preclinical models. Tumour Biol. 39(6):1010428317695528.

Lou JS et. al. (August 2016). Yu Ping Feng San reverses cisplatin-induced multi-drug resistance in lung cancer cells via regulating drug transporters and p62/TRAF6 signalling. Sci Rep. 6:31926.

Sarojini S et. al. (July 2015). A combination of high dose rate (10X FFF/2400 MU/min/10 MV X-rays) and total low dose (0.5 Gy) induces a higher rate of apoptosis in melanoma cells in vitro and superior preservation of normal melanocytes. Melanoma Res.

Sleire L et. al. (March 2015). Drug repurposing: sulfasalazine sensitizes gliomas to gamma knife radiosurgery by blocking cystine uptake through system Xc<sup>-</sup>, leading to glutathione depletion. Oncogene.

Błaszczak-Świątkiewicz K et. al. (March 2015). Some characteristics of activity of potential chemotherapeutics--benzimidazole derivatives. Adv Med Sci. 60(1):125-32.

Błaszczak-Świątkiewicz K et. al. (September 2014). Biological Evaluation of the Activity of Some Benzimidazole-4,7-dione Derivatives. Molecules. 19(10):15361-73.

Zhao J et. al. (August 2014). AKR1C3 Overexpression Mediates Methotrexate Resistance in Choriocarcinoma Cells. Int J Med Sci. 11(11):1089-97.

Gravina GL et. al. (June 2014). Torc1/Torc2 inhibitor, Palomid 529, enhances radiation response modulating CRM1-mediated survivin function and delaying DNA repair in prostate cancer models. Prostate. 74(8):852-68.

Błaszczak-Świątkiewicz K et. al. (February 2014). Biological approach of anticancer activity of new benzimidazole derivatives. Pharmacol Rep. 66(1):100-6.

Błaszczak-Świątkiewicz K et. al. (July 2013). Antiproliferative activity of new benzimidazole derivatives. Acta Biochim Pol. 60(3):427-33.

Takeda S et. al. (March 2013). (-)-Xanthatin up-regulation of the GADD45γ tumor suppressor gene in MDA-MB-231 breast cancer cells: role of topoisomerase IIα inhibition and reactive oxygen species. Toxicology. 305:1-9.

Hoeferlin LA et. al. (September 2011). Activation of p21-Dependent G1/G2 Arrest in the Absence of DNA Damage as an Antiapoptotic Response to Metabolic Stress. Genes Cancer. 2(9):889-99.

Yao JY et. al. (December 2010). TAp63 plays compensatory roles in p53-deficient cancer cells under genotoxic stress. Biochem Biophys Res Commun. 403(3-4):310-5.

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