Phospho-SMAD2 (Thr8) Recombinant Rabbit Monoclonal Antibody (11H10L30)/磷酸SMAD2（Thr8）重组兔单克隆抗体（11H10L30）

货号：700050

规格：100 µg

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：proprietary

物种：人

宿主：兔

抗体亚型：IgG

克隆号：11H10L30

荧光染料：其它

类型:	单抗	同型对照：
浓度：	0.5 mg/mL	用法：	3 µg(ChIP)；1 µg/mL(ICC)；1 µg/mL(IF)；Assay-Dependent(IP)；1-3 µg/mL(WB)

产品详细信息This antibody is predicted to react with bovine, Drosophila, mouse, rat and Xenopus based on sequence homology.Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones arescreened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.靶标信息SMAD2 (MADH2, MAD2) regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. SMAD2 interacts with the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation into the nucleus is a central event in TGF beta signaling. Phosphorylation of threonine 8 in the calmodulin-binding region of the MH1 domain by extracellular signal regulated kinase 1 (ERK1) enhances SMAD2 transcriptional activity, which is negatively regulated by calmodulin. In response to the TGF-beta signal, SMAD2 is phosphorylated by the TGF-beta receptors. The phosphorylation induces the dissociation of this protein with SARA and the association with the family member SMAD4. The association with SMAD4 is important for the translocation of this protein into the nucleus, where it binds to target promoters and forms a transcription repressor complex with other cofactors. SMAD2 can also be phosphorylated by activin type 1 receptor kinase, and mediates the signal from the activin. Alternatively spliced transcript variants encoding the SMAD2 protein have been observed.

数据：

Phospho-SMAD2 (Thr8) Antibody (700050) in IFImmunofluorescent analysis of SMAD2 (pT8) was done on 70% confluent log phase TGF-beta treated HeLa cells (serum starved for 16 hours followed by treatment with 20 ng/mL TGF-beta for 1 hour). The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with SMAD2 (pT8) Recombinant Rabbit Monoclonal Antibody (Product # 700050) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 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 Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing Nuclear localization of SMAD2 (pT8) and panel e is a no primary antibody control. The images were captured at 20X magnification. Phospho-SMAD2 (Thr8) Antibody (700050) in ChIPChromatin immunoprecipitation analysis of Phospho-SMAD2 (pThr8) was performed using cross-linked chromatin from 1 x 10^6 HCT116 human 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 Phospho-SMAD2 rabbit monoclonal antibody (Product # 700050). Chromatin aliquots from ~1 x 10^5 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 exon-1 or exon-2 of human Egr-1, or the imprinting control region (ICR) of the human H19 locus. 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 human Egr-1 and H19 loci are shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag lines represent introns, and the straight line represents upstream sequence. Regions amplified by Egr-1 and H19 primers are represented by black bars. Data courtesy of the Innovators Program.

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