GCN5 Monoclonal Antibody (2GC-2C11)/GCN5单克隆抗体（2GC-2C11）

货号：MA3-046

规格：50 µL

价格：4810

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Synthetic peptide coupled to ovalbumin correspondi

物种：人

宿主：小鼠

抗体亚型：其它

克隆号：2GC-2C11

荧光染料：其它

类型:	单抗	同型对照：
浓度：	Not Determined	用法：	1:100(ChIP)；1:2000(WB)

产品详细信息MA3-046 detects GCN5 in human samples.MA3-046 has been succesfully used in western blot and ChIP applications.Immunogen is a synthetic peptide corresponding to residues 118-132 of human GCN5靶标信息SAGA (Spt-Ada-Gen5 histone acetyltransferase complex) is a histoneacetylase complex which has Gcn5p as catalyst subunit and functions overlapping with the fundamental transcription factor TFIID which has Taf1p as catalyst subunit. However, SAGA and TFIID have different allotment and each accomplishes the important role in the transcription for Housekeeping gene group and Stress Responding gene group. Also, SAGA is a gigantic protein complex which is composed of Ada protein group (5 kinds), TBP related protein group (4 kinds), TAF protein group (5 kinds that also pertain to TFIID), and other protein groups (>6 kinds). As its molecular function, there is the chemical modification of histone, or the recruitment by direct interaction of transcription regulating factor on DNA, or the control of transcription starting reaction by TBP. Gcn5p that show histoneacetylase activity is one kind of the above-mentioned Ada protein group, and in the case of budding yeast it is composed of 439 amino acid residues.

数据：

GCN5 Antibody (MA3-046) in WBWestern blot analysis of GCN5 was performed by loading 10 µg of HeLa nuclear extract (lane 1), HeLa cytoplasmic extract (lane 2), U2OS nuclear extract (lane 3) and U2OS cytoplasmic extract (lane 4) 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 Gast Blotter (Product # 62288). Membrane was blocked in 5% Milk/TBST for one hour at room temperature. GCN5 was detected at approximately 94 kDa using GCN5 monoclonal antibody (Product # MA3-046) at a dilution of 1:2000 overnight at 4°C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:5,000 for one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076) and the myECL Imager (Product # 62236). GCN5 Antibody (MA3-046) in ChIPChromatin immunoprecipitation analysis of HIV1 GCN5 was performed using cross-linked chromatin from Human 5A8 J-lat T lymphocytes culture latently infected with HIV1 and treated with 10 µg/mL PHA (phytohemaglutin) for 0, 4, and 8 hours. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of GCN5 monoclonal antibody (Product # MA3-046). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers shown to amplify exon-1 of the Egr1 gene or HIV gag gene. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. Schematic representations of the Egr1 and HIV1 gag 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 the Innovators Program.

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