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EpiQuik Tissue Chromatin Immunoprecipitation (ChIP) Kit

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For immunoprecipitating chromatin specifically from tissue input samples via microplate format

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Suggested Workflow
Chromatin Isolation
 
 
Chromatin Shearing
 
 
ChIP
 
 
PCR Analysis
 
Schematic procedure of the EpiQuik™ Tissue Chromatin Immunoprecipitation (ChIP) Kit.Fig. 2. Comparative overview of the EpiQuik™ Tissue Chromatin Immunoprecipitation (ChIP) Kit.
Input Type: Chromatin
Research Area: Chromatin & Transcription
Target Application: Immunoprecipitation
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-2003-124 reactions $234.00 
P-2003-248 reactions $380.00 
P-2003-396 reactions $559.00 
Availability: Usually Ships in 1 Day or Same day delivery Same Day NY Delivery 
Product Overview

The EpiQuik™ Tissue Chromatin Immunoprecipitation (ChIP) Kit is a convenient package of tools that allows the experimenter to quickly perform chromatin immunoprecipitation (ChIP) from mammalian tissue. The kit is ready-to-use and provides all the essential components needed to carry out a successful ChIP experiment on tissue. The EpiQuik™ ChIP kits are suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, ChIP-Seq, and ChIP-on-chip. This kit has the following advantages:

  • The fastest procedure available, which can be finished within 5 hours.
  • The ChIP procedure has been drastically simplified -- extremely short and easy to follow.
  • Strip microplate format makes the assay flexible: manual or high throughput.
  • Columns for DNA purification are included, which save time and reduce labor.
  • Compatible with all DNA amplification-based approaches.
  • Achieves very reliable and consistent assay conditions.

See also a quick chart to compare ChIP kits.

Background Information
Protein-DNA interaction play a critical role for cellular functions such as signal transduction, gene transcription, chromosome segregation, DNA replication and recombination, and epigenetic silencing. Identifying the genetic targets of DNA binding proteins and knowing the mechanisms of protein-DNA interaction is important for understanding cellular process. Chromatin Immunoprecipitation (ChIP) offers an advantageous tool for studying protein-DNA interactions. Unlike other methods such as EMASA, DNA microarrays, and report gene assays, which analyze direct interactions between protein and DNA in vitro, ChIP can detect that a specific protein binds to the specific sequences of a gene in living cells.   

Principle & Procedure
This ChIP kit includes all reagents required for carrying out a successful chromatin immunoprecipitation from mammalian tissue. Particularly, this kit includes a positive control antibody (RNA polymerase II), a negative control normal mouse IgG, and GAPDH primers that can be used as a positive control to demonstrate the efficacy of the kit reagents and protocol. RNA polymerase II is considered to be enriched in the GAPDH gene promoter that is expected to be undergoing transcription in most growing mammalian cells and can be immunoprecipitated by RNA polymerase II but not by normal mouse IgG. In this ChIP, cells are cross-linked with formaldehyde and chromatin is extracted. The chromatin is then sheared and added into the microwell immobilized with affinity antibodies. Cross-linked DNA is released from antibody-captured protein-DNA complex, reversed and purified through the specifically designed F-Spin Column. Eluted DNA can be used for various down-stream applications. 

Starting Materials
Starting materials can include various tissue samples. In general, the input amount should be from 10 to 30 mg of tissue for each reaction.


Fig. 1. Schematic procedure of the EpiQuik™ Tissue Chromatin Immunoprecipitation (ChIP) Kit.


Fig. 2. Comparative overview of the EpiQuik™ Tissue Chromatin Immunoprecipitation (ChIP) Kit.

Product Components

CP1 (Wash Buffer)
CP2 (Antibody Buffer)
CP3 (Lysis Buffer)
CP4 (ChIP Dilution Buffer)
CP5 (DNA Release Buffer)
CP6 (Reverse Buffer)
CP7 (Binding Buffer)
CP8 (Elution Buffer)
Homogenizing Buffer
Protease Inhibitor Cocktail (100X)*
Normal Mouse IgG (1 mg/ml)*
Anti-RNA Polymerase II (1 mg/ml)*
Proteinase K (10 mg/ml)*
Control Primer (GAPDH)
Forward (20 µM)*
Reverse (20 µM)*
8-Well Assay Strips (with Frame)
8-Well Strip Caps
F-Spin Column
F-Collection Tube
User Guide

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

Frequently Asked Q's

1. Can IP be overnight?
Yes.

2. Is there any background DNA by mouse IgG IP?
Yes, a little < 2 ng.

3. Can any species Ab be used for the IP?
Yes, if it is an IgG antibody.

4. Can kit P-2003 be used for frozen tissues?
Yes, but it is not as good as fresh samples.

5. How much volume of elution should be added into PCR reaction?
2 µl.

6. Are cell extracts containing 1%SDS compatible with CP4?
No, the SDS concentration in IP solution should be less than 0.1%.

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.

[Material Safety Data Sheet]
Product Citations

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Heesch MW et. al. (July 2016). Transcriptional control, but not sub-cellular location, of PGC-1α is altered following exercise in a hot environment. J Appl Physiol (1985). :jap.01065.2015.

Chater-Diehl EJ et. al. (May 2016). Alteration of Gene Expression, DNA Methylation, and Histone Methylation in Free Radical Scavenging Networks in Adult Mouse Hippocampus following Fetal Alcohol Exposure. PLoS One. 11(5):e0154836.

Zhang X et. al. (January 2016). Autosomal gsdf acts as a male sex initiator in the fish medaka. Sci Rep. 6:19738.

Wiegreffe C et. al. (July 2015). Bcl11a (Ctip1) Controls Migration of Cortical Projection Neurons through Regulation of Sema3c. Neuron. 87(2):311-25.

Li D et. al. (June 2015). Epigenetic regulation of traf2- and Nck-interacting kinase (TNIK) in polycystic ovary syndrome. Am J Transl Res.. 7(6):1152-60.

Tu P et. al. (March 2015). Liver histone H3 methylation and acetylation may associate with type 2 diabetes development. J Physiol Biochem. 71(1):89-98.

Wang Y et. al. (January 2015). Novel curcumin analog C66 prevents diabetic nephropathy via JNK pathway with the involvement of p300/CBP-mediated histone acetylation. Biochim Biophys Acta. 1852(1):34-46.

Angrisano T et. al. (September 2014). Epigenetic switch at atp2a2 and myh7 gene promoters in pressure overload-induced heart failure. PLoS One. 9(9):e106024.

Shen W et. al. (June 2014). Epigenetic modification of the leptin promoter in diet-induced obese mice and the effects of N-3 polyunsaturated fatty acids. Sci Rep. 4:5282.

Zhang W et. al. (May 2014). Prenatal alcohol exposure causes the over-expression of DHAND and EHAND by increasing histone H3K14 acetylation in C57 BL/6 mice. Toxicol Lett. 228(3):140-146.

Kuroda N et. al. (April 2014). Apoptotic response through a high mobility box 1 protein-dependent mechanism in LPS/GalN-induced mouse liver failure and glycyrrhizin-mediated inhibition. PLoS One. 9(4):e92884.

Monteiro JB et. al. (March 2014). Endometriosis is characterized by a distinct pattern of histone 3 and histone 4 lysine modifications. Reprod Sci. 21(3):305-18.

Anier K et. al. (October 2013). S-adenosylmethionine modifies cocaine-induced DNA methylation and increases locomotor sensitization in mice. Int J Neuropsychopharmacol. 16(9):2053-66.

Das B et. al. (May 2013). The sld genetic defect: two intronic CA repeats promote insertion of the subsequent intron and mRNA decay. J Biol Chem. 288(21):14742-55.

Fan-xin M et. al. (June 2012). Heat shock factor 1 regulates the expression of the TRPV1 gene in the rat preoptic-anterior hypothalamus area during lipopolysaccharide-induced fever. Exp Physiol. 97(6):730-40.

Sui L et. al. (May 2012). Epigenetic regulation of reelin and brain-derived neurotrophic factor genes in long-term potentiation in rat medial prefrontal cortex. Neurobiol Learn Mem. 97(4):425-40.

John A et. al. (May 2012). Bcl11a is required for neuronal morphogenesis and sensory circuit formation in dorsal spinal cord development. Development. 139(10):1831-41.

Wu CH et. al. (November 2011). A double-edged sword role for ubiquitin-proteasome system in brain stem cardiovascular regulation during experimental brain death. PLoS One. 6(11):e27404.

Sui L et. al. (December 2010). Effects of perinatal hypothyroidism on regulation of reelin and brain-derived neurotrophic factor gene expression in rat hippocampus: Role of DNA methylation and histone acetylation. Steroids. 75(12):988-97.

Aoyama T et. al. (September 2010). Histone modifiers, YY1 and p300, regulate the expression of cartilage-specific gene, chondromodulin-I, in mesenchymal stem cells. J Biol Chem. 285(39):29842-50.

Heinig M et. al. (September 2010). A trans-acting locus regulates an anti-viral expression network and type 1 diabetes risk. Nature. 467(7314):460-4.

Aoki T et. al. (September 2010). Ets-1 promotes the progression of cerebral aneurysm by inducing the expression of MCP-1 in vascular smooth muscle cells. Gene Ther. 17(9):1117-23.

Chung S et. al. (August 2010). Glutaredoxin 1 regulates cigarette smoke-mediated lung inflammation through differential modulation of I{kappa}B kinases in mice: impact on histone acetylation. Am J Physiol Lung Cell Mol Physiol. 299(2):L192-203.

Yamaguchi T et. al. (August 2010). Cortisol is involved in temperature-dependent sex determination in the Japanese flounder. Endocrinology. 151(8):3900-8.

Endo T et. al. (June 2010). Runx2 deficiency in mice causes decreased thyroglobulin expression and hypothyroidism. Mol Endocrinol. 24(6):1267-73.

Kanfi Y et. al. (April 2010). SIRT6 protects against pathological damage caused by diet-induced obesity. Aging Cell. 9(2):162-73.

Molteni R et. al. (March 2010). Reduced function of the serotonin transporter is associated with decreased expression of BDNF in rodents as well as in humans. Neurobiol Dis. 37(3):747-55.

Herpin A et. al. (February 2010). Transcriptional rewiring of the sex determining dmrt1 gene duplicate by transposable elements. PLoS Genet. 6(2):e1000844.

Joh K et. al. (December 2009). Antisense transcription occurs at the promoter of a mouse imprinted gene, commd1, on the repressed paternal allele. J Biochem. 146(6):771-4.

Yue L et. al. (April 2009). Peroxisome proliferator-activated receptor {gamma} stimulation of adipocyte ApoE gene transcription mediated by the liver receptor X pathway. J Biol Chem. 284(16):10453-61.

Doonan F et. al. (April 2009). Rosiglitazone acts as a neuroprotectant in retinal cells via up-regulation of sestrin-1 and SOD-2. J Neurochem. 109(2):631-43.

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