相关类别
一般描述
The ChIPAb+ HDAC1 set includes the HDAC1 antibody, the negative control antibody supernatant (mouse IgG), and qPCR primers flanking an Sp1 binding site in human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The HDAC1 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of HDAC1 associated chromatin.
免疫原
应用
Sonicated chromatin prepared from untreated or actinomycin D treated (30 ng/mL, 6 hrs.) U2OS cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 4 μL of Mouse Anti-HDAC1 and the Magna ChIP® G (Cat. # 17-611) Kit. Immunoprecipitation of HDAC1 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Decrease is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. As previously published, HDAC1 association with this region of the p21 promoter decreases upon actinomycin D treatment.
Western Blot Analysis:
3T3/A31 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-HDAC1 (1:2500). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Epigenetics & Nuclear Function
Chromatin Biology
生化/生理作用
包装
外形
Negative ChIP Control Supernatant. One vial containing 40 uL of mouse IgG containing supernatant with 0.05% sodium azide. Store at -20°C.
ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter. Store at -20°C.
P21 Forward: CCC ACA GCA GAG GAG AAA GAA
P21 Reverse: CTG GAA ATC TCT GCC CAG ACA
制备说明
分析说明
Included negative control antibody supernatant mouse IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.
法律信息
免责声明
储存分类代码
10 - Combustible liquids
相关内容
"Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms instead of by alterations in DNA sequence. These changes can be cell- or tissue-specific, and can be passed on to multiple generations. Epigenetic regulation enriches DNAbased information, allowing a cell to vary its response across diverse biological and environmental contexts. Although epigenetic mechanisms are primarily centered in the nucleus, these mechanisms can be induced by environmental signals such as hormones, nutrients, stress, and cellular damage, pointing to the involvement of cytoplasmic and extracellular factors in epigenetic regulation."
Epigenetics describes heritable changes in gene expression caused by non-genetic mechanisms. Epigenetic regulation allows a cell to vary its response based on its biological and environmental contexts. Epigenetic changes can effect transcriptional and post-transcriptional regulation via mechanisms such as histone modification, chromatin and nucleosome remodeling, DNA methylation, and small and non-coding RNA-mediated regulation. These mechanisms, in cooperation with transcription factors and other nucleic acid-binding proteins, regulate gene expression. Epigenetic mechanisms of gene regulation impacts diverse areas of research—from agriculture to human health. Common epigenetic assays such as chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) rely on high quality antibodies that recognize specific epigenetic modifications for accurate results. EMD Millipore offers over 100 ChIPAb+™ and RIPAb+™ validated antibody kits that are quality tested on ChIP/RIP assays and are conveniently provided with control qPCR primers and negative control antibodies to ensure first time ChIP/RIP success.
Signaling Product Guide: Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.
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