Phospho-p70 S6 Kinase (Thr421, Ser424) Recombinant Rabbit Monoclonal Antibody (B12H16L8)/Phospho-p70 S6激酶（Thr421，Ser424）重组兔单克隆抗体（B12H16L8）

货号：701083

规格：100 µg

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Phosphopeptide corresponding to amino acids 417-42

物种：其它

宿主：大鼠

抗体亚型：IgG

克隆号：B12H16L8

荧光染料：其它

类型:	单抗	同型对照：
浓度：	0.5 mg/mL	用法：	1 µL(ChIP)；1:500-1:5000(WB)

产品详细信息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 are screened 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The p70 S6 kinase (p70 Ribosomal Protein S6 Kinase, p70S6K) is a 70 kDa member of the ribosomal S6 kinase (RSK) family of serine/threonine kinases. p70 S6 kinase is predominantly localized in the cytoplasm, and is essential in growth factors regulated cell proliferation, pathways involving cell motility, such as metastases, the immune response, and tissue repair. p70 S6 kinase acts downstream of phosphoinositide (PI) 3-kinase, and its main physiological target is the S6 ribosomal protein, whichis involved in upregulation of protein synthesis. The kinase activity of p70 S6 kinase leads to an increase in protein synthesis and cell proliferation. Amplification of the region of DNA encoding p70 S6 kinase and overexpression are seen in some breast cancer cell lines.

数据：

Phospho-p70 S6 Kinase (Thr421, Ser424) Antibody (701083) in WBWestern blot analysis of p70S6K (pTpS421/424) was performed by loading 30 µg of untreated HeLa cells (lane 1), HeLa cells treated with IGF1 (150 ng/mL for 15 minutes) (lane 2) and U-87 MG lysates (lane 3) using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), Xcell SureLock Electrophoresis system (Product # EI0002), Novex sharp Pre-stained Protein Standard (Product # LC5800). Proteins were transferred to a PVDF membrane and blocked with 5% skim milk for 1 hour at room temperature. p70S6K (pTpS421/424) was detected at ~70 kDa using p70S6K (pTpS421/424) Recombinant Rabbit Monoclonal Antibody (Product # 701083) at a 1:1000 dilution in 2.5% skim milk at 4° C overnight on a rocking platform. To confirm specificity, competition was performed with the phosphopeptide (10 µg/mL) (as shown in the corresponding blot on the right). Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106). Phospho-p70 S6 Kinase (Thr421, Ser424) Antibody (701083) in ChIPChromatin immunoprecipitation analysis of Phospho-p70 S6 Kinase (pThr421+pSer424) was performed using cross-linked chromatin from 1 x 10^6 HCT116 human colon carcinoma cells treated with serum for 0, 15, and 60 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-p70 S6 Kinase (pThr421+pSer424) rabbit monoclonal antibody (Product # 701083). 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 -3.2kb upstream of the human FOS gene, or exon-4 of human FOS. 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 FOS locus is shown above the data where boxes represent exons (grey boxes = translated regions, white boxes = untranslated regions), the zigzag lines represent introns, and the straight line represents upstream sequence. Regions amplified by FOS primers are represented by black bars. Data courtesy of the Innovators Program.

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