p300 Polyclonal Antibody/p300多克隆抗体

货号：PA1-848

规格：100 µg

价格：5468

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Synthetic peptide corresponding to residues G(139)

物种：人

宿主：兔

抗体亚型：IgG

荧光染料：其它

类型:	多抗	同型对照：
浓度：	1 mg/mL	用法：	1-3 µL(ChIP)；5 µg/mL(ICC)；5 µg/mL(IF)；1-5 µg/mL(WB)

产品详细信息PA1-848 detects p300 from human cells.PA1-848 has been successfully used in Western blot and ChIP procedures. By Western blot, this antibody detects an ~265 kDa protein representing p300 from HeLa cell lysate.The PA1-848 immunogen is a synthetic peptide corresponding to residues G(139) T S G P N Q G P T Q S T(151) C in the nuclear factor binding domain of human p300. The PA1-848 immunizing peptide (Cat. # PEP-053) is available for use in neutralization and control experiments.靶标信息EP300 encodes the adenovirus E1A-associated cellular p300 transcriptional co-activator protein. p300 is related by sequence to CPB (CREB-binding protein [CREB: cyclic-AMP responsive element binding protein]), and like CPB can stimulate transcription through activation of CREB. This EP300 activity is specifically inhibited by the adenovirus oncoprotein E1A. EP300 has also been identified as a co-activator of HIF1A (hypoxia-inducible factor 1 alpha), and thus plays a role in the stimulation of hypoxia-induced genes such as VEGF.

数据：

p300 Antibody (PA1-848) in IFImmunofluorescence analysis of p300 was performed using 70% confluent serum starved and released HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with p300 Rabbit polyclonal antibody (Product # PA1-848) at 5 µg/mL in 0.1% BSA, incubated overnight at 4 degree Celsius and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents the untreated cells showing cytoplasmic localization of p300. Panel f shows control cells with no primary antibody to assess background. The images were captured at 60X magnification. p300 Antibody (PA1-848) in ChIPChromatin immunoprecipitation analysis of p300 was performed using cross-linked chromatin from 1x10^6 HCT116 colon carcinoma cells treated with serum for 0, 15, and 30 minutes. 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 a p300 polyclonal antibody (Product # PA1-848). Chromatin aliquots from ~1x10^5 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 to amplify -15kb upstream of the Egr1 gene or exon-1 or exon-2 of Egr1. 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. A schematic representation of the Egr-1 locus is 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 Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.

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参考文献：

1. Nucleic acids researchDYRK1A interacts with histone acetyl transferase p300 and CBP and localizes to enhancers.2. Molecular endocrinology (Baltimore, Md.)Research resource: identification of novel coregulators specific for thyroid hormone receptor-β2.3. The Journal of biological chemistryG Protein-coupled Receptor 40 (GPR40) and Peroxisome Proliferator-activated Receptor γ (PPARγ): AN INTEGRATED TWO-RECEPTOR SIGNALING PATHWAY.

技术参数

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