H3K9me2S10phK27me2S28ph Monoclonal Antibody (51TA-2H12)/H3K9me2S10phK27me2S28ph单克隆抗体（51TA-2H12）

货号：MA3-065

规格：50 µL

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Ovalbumin conjugated synthetic peptide (ASGSSRSKS[

物种：人

宿主：小鼠

抗体亚型：其它

克隆号：51TA-2H12

荧光染料：其它

类型:	单抗	同型对照：
浓度：	Conc. Not Determined	用法：	Assay dependent(ChIP)；1-150(IP)；1-2000(WB)

产品详细信息MA3-065 detects dimethyl-phospho-histone H3 Lys9/Lys27 pSer10/pSer28 in human samples and has successfully been used in western blot and immunoprecipitation applications.The MA3-065 immunogen is a synthetic peptide conjugated to ovalbumin corresponding to residues 180-195 of human TAF10 and only recognizes phosphorlayted pSer10 or pSer28 of Histone H3 when the adjacent Lys is dimethylated.靶标信息Histone H3 is one of the DNA-binding proteins found in the chromatin of all eukaryotic cells. H3 along with four core histone proteins binds to DNA forming the structure of the nucleosome. Histones play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. Post translationally, histones are modified in a variety of ways to either directly change the chromatin structure or allow for the binding of specific transcription factors. The N-terminal tail ofhistone H3 protrudes from the globular nucleosome core and can undergo several different types of post-translational modification that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.

数据：

H3K9me2S10phK27me2S28ph Antibody (MA3-065) in WBWestern blot analysis of dimethyl-phospho-Histone H3 Lys9/27 pSer10/28 was performed by loading 10 µg of acid extracted HeLa nuclear lysate extracted from cells not treated (lane 1) or treated with 100 nM calyculin A (Product # PHZ1044) (lane 2) for 30 min in reducing sample buffer (Product # 39000) and Page Ruler Plus Protein Ladder (Product # 26619) onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4201BX10). Proteins were transferred to nitrocellulose membrane (Product # 88018) with Transfer Buffer (Product # 84731) using the G2 Fast Blotter (Product # 62288). Membrane was blocked in StartingBlock T20 (Product # 37543) for 30 min at room temperature. Dimethyl-phospho-Histone H3 Lys9/27 pSer10/28 was detected at approximately 17 kDa using an H3K9/27me2S10/28ph monoclonal antibody (Product # MA3-065) at a dilution of 1:2000 in StartingBlock T20 overnight at at 4°C on a rocking platform, followed by a goat anti-mouse superclonal IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:5000 for one hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product # 62236). H3K9me2S10phK27me2S28ph Antibody (MA3-065) in ChIPMultiplex microplate Matrix ChIP has been described in detail (http://www.ncbi.nlm.nih.gov/pubmed/25959381). Briefly, frozen mouse lung tissue fragments (10 to 20mg) were cross-linked with formaldehyde and chromatin was sheared using a Bioruptor. ChIP assays were performed using protein A-coated 96-well polypropylene microplates with 1uL/100uL well volume of H3K9/27me2S10/28ph monoclonal antibody (Product # MA3-065). Quantitative real-time PCRs were performed in quadruplicate using 1 to 2uL of DNA with primers to exon 1 of Alb (a gene repressed in the lung) and exon 6 of Sftpc (a gene normally expressed in the lung). PCR calibration curves were generated for each primer pair from a dilution series of total mouse genomic DNA. The PCR primer efficiency curve was fit to cycle threshold (Ct) versus log (genomic DNA concentration) by using an r2 best fit. DNA concentration values for each ChIP and input DNA sample were calculated from their respective average Ct values. Final results are expressed as fraction of input DNA. Schematic representations of the Alb and Sftpc locus are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by the primers are represented by black bars. Data courtesy of Dr. Bomsztyk’s lab.

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