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Product listing: Sunitinib #12328 to SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003

Molecular Weight:616.84 g/mol

Background: TPA (12-O-tetradecanoylphorbol-13-acetate) / PMA (phorbol-12-myristate-13-acetate) is the most commonly used phorbol ester. It binds and activates protein kinase C, causing a wide range of effects in cells and tissues (1,2). TPA has been demonstrated to be a potent tumor promoter in mouse skin (3). Conversely, it has been shown to be an effective cancer therapeutic agent in myelocytic leukemia patients, and has been indicated as a potential colorectal cancer therapeutic (4-5). TPA has also been shown to increase white blood cell and neutrophil counts in solid tumor cancer patients (6).

Molecular Weight:302.4 g/mol

Background: Trichostatin A is an antifungal antibiotic derived from Streptomyces that inhibits mammalian histone deacetylases (1,2). When used at nanomolar concentrations, trichostatin A causes the accumulation of acetylated histones in a quite specific and reversible manner (2,3). Trichostatin A has also been shown to inhibit both G1- and G2-phases of the mammalian cell cycle and has been tested for use as a potential anticancer agent (2-5).

Molecular Weight:315.76 g/mol

Background: Tyrphostin AG 1478 is a tyrosine kinase inhibitor specifically selective to EGFR (ErbB1), with an IC50 of about 3 nM in vitro (1,2). Treatment of cell with 50-150 nM of AG 1478 can substantially block EGFR activiation in vivo (3). In addition to EGFR, AG 1478 also inhibits ErbB4 activation induced by radiation in cancer cells (4). Testing of AG 1478 alone or in combination with other treatments to assess anti-tumor and anti-fibrotic effectiveness has yielded promising results (5-8).

Molecular Weight:380.50 g/mol

Background: MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221, located in the activation loop of subdomain VIII, by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines and also by membrane depolarization and calcium influx (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC-12 cells (4). MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII.

Molecular Weight:428.4 g/mol

Background: Wortmannin is a very potent, specific and direct inhibitor of PI3 kinase, originally derived from fungus (1,2). The inhibition is irreversible and noncompetitive. Wortmannin does not inhibit PI4 kinase, protein kinase C or protein tyrosine kinase (3).

$714
24 assays
1 Kit
Next generation sequencing (NG-seq) is a high throughput method that can be used downstream of chromatin immunoprecipitation (ChIP) assays to identify and quantify target DNA enrichment across the entire genome. The SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® contains all of the enzymes and buffers necessary to generate high quality DNA sequencing libraries from ChIP DNA for next-generation sequencing on the Illumina® platform. The fast, user-friendly workflow minimizes hands-on time needed for generation and purification of DNA libraries. This product must be used in combination with SimpleChIP® ChIP-seq Multiplex Oligos for Illumina® (Single Index Primers) #29580 or SimpleChIP® ChIP-seq Multiplex Oligos for Illumina® (Dual Index Primers) #47538.This product provides sufficient amounts of reagents for 24 reactions and is compatible with both enzymatic- and sonication-fragmented, ChIP-enriched DNA. This product is compatible with SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003, SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005, and SimpleChIP® Plus Sonication Chromatin IP Kit #56383. This product is not compatible with SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads) #9002 and SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads) #9004 because agarose beads are blocked with sonicated salmon sperm DNA, which will contaminate DNA library preps and NG-seq.
REACTIVITY
All Species Expected

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to identify multiple proteins associated with a specific region of the genome, or the opposite, to identify the many regions of the genome bound by a particular protein (3-6). It can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors and DNA repair proteins. When performing the ChIP assay, cells or tissues are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. The chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or Quantitative Real-Time PCR can be used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing, or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8).

$561
96 assays
1 Kit
Next generation sequencing (NG-seq) is a high throughput method that can be used downstream of chromatin immunoprecipitation (ChIP) assays to identify and quantify target DNA enrichment across the entire genome. SimpleChIP® ChIP-seq Multiplex Oligos for Illumina® (Dual Index Primers) contains adaptors and primers that are ideally suited for multiplex sample preparation for NG-seq on the Illumina® platform. This kit can be used to generate up to 96 distinct, barcoded ChIP-seq DNA libraries that can be combined into a single sequencing reaction. This product provides enough reagents to support up to 96 DNA sequencing libraries, and must be used in combination with SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795.This product is compatible with SimpleChIP® Enzymatic ChIP Kit (Magnetic Beads) #9003, SimpleChIP® Plus Enzymatic ChIP Kit (Magnetic Beads) #9005, and SimpleChIP® Plus Sonication ChIP kit #56383. This product is not compatible with SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads) #9002 and SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads) #9004 because agarose beads are blocked with sonicated salmon sperm DNA, which will contaminate DNA library preps and NG-seq.
REACTIVITY
All Species Expected

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to identify multiple proteins associated with a specific region of the genome, or the opposite, to identify the many regions of the genome bound by a particular protein (3-6). It can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors and DNA repair proteins. When performing the ChIP assay, cells or tissues are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. The chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or Quantitative Real-Time PCR can be used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing, or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8).

$184
24 assays
1 Kit
Next generation sequencing (NG-seq) is a high throughput method that can be used downstream of chromatin immunoprecipitation (ChIP) assays to identify and quantify target DNA enrichment across the entire genome. SimpleChIP® ChIP-seq Multiplex Oligos for Illumina® (Single Index Primers) contains adaptors and primers that are ideally suited for multiplex sample preparation for NG-seq on the Illumina® platform. This kit can be used to generate up to 12 distinct, barcoded ChIP-seq DNA libraries that can be combined into a single sequencing reaction.This product provides enough reagents to support up to 24 DNA sequencing libraries, and must be used in combination with the SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795.This product is compatible with SimpleChIP® Enzymatic ChIP Kit (Magnetic Beads) #9003, SimpleChIP® Plus Enzymatic ChIP Kit (Magnetic Beads) #9005, and SimpleChIP® Plus Sonication ChIP kit #56383. This product is not compatible with SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads) #9002 and SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads) #9004 because agarose beads are blocked with sonicated salmon sperm DNA, which will contaminate DNA library preps and NG-seq.
REACTIVITY
All Species Expected

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to identify multiple proteins associated with a specific region of the genome, or the opposite, to identify the many regions of the genome bound by a particular protein (3-6). It can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors and DNA repair proteins. When performing the ChIP assay, cells or tissues are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. The chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or Quantitative Real-Time PCR can be used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing, or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8).

$76
30 immunoprecipitations
1 Kit
This product is offered to conveniently provide additional ChIP buffer reagents for preparing, immunoprecipitating, washing, and eluting chromatin using our SimpleChIP® (#9002, #9003) and SimpleChIP® Plus (#9004, #9005) Enzymatic Chromatin IP Kits, as well as our SimpleChIP® Sonication Chromatin IP kit (#56383). These SimpleChIP® kits provide all the reagents required for performing the recommended of chromatin preparations (or optimizations) and chromatin immunoprecipitation (ChIP) assays, however there are instances where extra ChIP Buffer, ChIP Elution buffer, and NaCl are desired.
REACTIVITY
All Species Expected
$107
36 DNA Purifications
1 Kit
This product is offered to conveniently provide additional ChIP buffer reagents for preparing, immunoprecipitating, washing, and eluting chromatin using our SimpleChIP® (#9002, #9003) and SimpleChIP® Plus (#9004, #9005) Enzymatic Chromatin IP Kits, as well as our SimpleChIP® Sonication Chromatin IP kit (#56383). These SimpleChIP® kits provide all the reagents required for performing the recommended of chromatin preparations (or optimizations) and chromatin immunoprecipitation (ChIP) assays, however there are instances where extra ChIP Buffer, ChIP Elution buffer, and NaCl are desired.The DNA purification columns combine the convenience of spin-column technology with the selective binding properties of a uniquely designed silica membrane that allows for efficient recovery of DNA and removal of protein contaminants without the need for phenol/chloroform extractions and ethanol precipitations. After DNA purification, the enrichment of particular DNA sequences can be analyzed by a variety of methods.
$76
30 immunoprecipitations
1 Kit
This product is offered to conveniently provide additional Buffer A and Buffer B reagents for harvesting cells and tissues and preparing chromatin using our SimpleChIP® (#9002, #9003) and SimpleChIP® Plus (#9004, #9005) Enzymatic Chromatin IP Kits. These kits provide all the reagents required for performing 6 chromatin preparations (or optimizations) and 30 chromatin immunoprecipitation (ChIP) assays, however there are instances where extra Buffer A and Buffer B reagents are desired.
$490
30 immunoprecipitations
1 Kit
The SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads) #9002 contains the buffers and reagents necessary to perform up to 6 chromatin preparations and 30 chromatin immunoprecipitations and is optimized for 4 X 107 cells per experiment. A complete assay can be performed in as little as two days and can easily be scaled up or down for use with more or fewer cells.Cells are fixed with formaldehyde and lysed, and chromatin is fragmented by partial digestion with Micrococcal Nuclease to obtain chromatin fragments of 1 to 5 nucleosomes. Enzymatic fragmentation of chromatin is much milder than sonication and eliminates problems resulting from variability in sonication power and emulsification of chromatin during sonication, which can result in incomplete fragmentation of chromatin or loss of antibody epitopes due to protein denaturation and degradation. Chromatin immunoprecipitations are performed using ChIP-validated antibodies and ChIP-Grade Protein G Agarose Beads. After reversal of protein-DNA cross-links, the DNA is purified using DNA purification spin columns, allowing for easy and efficient recovery of DNA and removal of protein contaminants without the need for phenol/chloroform extractions and ethanol precipitations. The enrichment of particular DNA sequences during immunoprecipitation can be analyzed by a variety of methods, including standard PCR and quantitative real-time PCR. Please note that this kit is not compatible with ChIP-seq because the ChIP-Grade Protein G Agarose Beads are blocked with sonicated salmon sperm DNA, which interferes with downstream sequencing.The SimpleChIP® Kit also provides important controls to ensure a successful ChIP experiment. The kit contains a positive control Histone H3 Antibody, a negative control Normal Rabbit IgG Antibody and primer sets for PCR detection of the human and mouse ribosomal protein L30 (RPL30) genes. Histone H3 is a core component of chromatin and is bound to most DNA sequences throughout the genome, including the RPL30 locus. Thus, the Histone H3 Antibody provides a universal positive control that should enrich for almost any locus examined.

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to identify multiple proteins associated with a specific region of the genome, or the opposite, to identify the many regions of the genome bound by a particular protein (3-6). It can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors and DNA repair proteins. When performing the ChIP assay, cells or tissues are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. The chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or Quantitative Real-Time PCR can be used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing, or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8).

$504
30 immunoprecipitations
1 Kit
The SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 contains the buffers and reagents necessary to perform up to 6 chromatin preparations and 30 chromatin immunoprecipitations and is optimized for 4 X 107 cells per experiment. A complete assay can be performed in as little as two days and can easily be scaled up or down for use with more or fewer cells.Cells are fixed with formaldehyde and lysed, and chromatin is fragmented by partial digestion with Micrococcal Nuclease to obtain chromatin fragments of 1 to 5 nucleosomes. Enzymatic fragmentation of chromatin is much milder than sonication and eliminates problems resulting from variability in sonication power and emulsification of chromatin during sonication, which can result in incomplete fragmentation of chromatin or loss of antibody epitopes due to protein denaturation and degradation. Chromatin immunoprecipitations are performed using ChIP-validated antibodies and ChIP-Grade Protein G Magnetic Beads. After reversal of protein-DNA cross-links, the DNA is purified using DNA purification spin columns, allowing for easy and efficient recovery of DNA and removal of protein contaminants without the need for phenol/chloroform extractions and ethanol precipitations. The enrichment of particular DNA sequences during immunoprecipitation can be analyzed by a variety of methods, including standard PCR, quantitative real-time PCR, or amplification for ChIP on chip, sequencing or cloning techniques. This kit is compatible with ChIP-seq.The SimpleChIP® Kit also provides important controls to ensure a successful ChIP experiment. The kit contains a positive control Histone H3 Antibody, a negative control Normal Rabbit IgG Antibody and primer sets for PCR detection of the human and mouse ribosomal protein L30 (RPL30) genes. Histone H3 is a core component of chromatin and is bound to most DNA sequences throughout the genome, including the RPL30 locus. Thus, the Histone H3 Antibody provides a universal positive control that should enrich for almost any locus examined.

Background: The chromatin immunoprecipitation (ChIP) assay is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1,2). This assay can be used to identify multiple proteins associated with a specific region of the genome, or the opposite, to identify the many regions of the genome bound by a particular protein (3-6). It can be used to determine the specific order of recruitment of various proteins to a gene promoter or to "measure" the relative amount of a particular histone modification across an entire gene locus (3,4). In addition to histone proteins, the ChIP assay can be used to analyze binding of transcription factors and co-factors, DNA replication factors and DNA repair proteins. When performing the ChIP assay, cells or tissues are first fixed with formaldehyde, a reversible protein-DNA cross-linking agent that "preserves" the protein-DNA interactions occurring in the cell (1,2). Cells are lysed and chromatin is harvested and fragmented using either sonication or enzymatic digestion. The chromatin is then immunoprecipitated with antibodies specific to a particular protein or histone modification. Any DNA sequences that are associated with the protein or histone modification of interest will co-precipitate as part of the cross-linked chromatin complex and the relative amount of that DNA sequence will be enriched by the immunoselection process. After immunoprecipitation, the protein-DNA cross-links are reversed and the DNA is purified. Standard PCR or Quantitative Real-Time PCR can be used to measure the amount of enrichment of a particular DNA sequence by a protein-specific immunoprecipitation (1,2). Alternatively, the ChIP assay can be combined with genomic tiling micro-array (ChIP on chip) techniques, high throughput sequencing, or cloning strategies, all of which allow for genome-wide analysis of protein-DNA interactions and histone modifications (5-8).