Phospho-Histone H3 (Ser10) Recombinant Rabbit Monoclonal Antibody (9H12L10)/磷酸化组蛋白H3（Ser10）重组兔单克隆抗体（9H12L10）

货号：701258

规格：100 µg

价格：4809

产品类型：抗体和染料

品牌：Thermo Fisher

抗原：Phosopeptide corresponding to amino acids 4–16 of human Histone H3 [pS 10]

物种：人

宿主：兔

抗体亚型：IgG

克隆号：9H12L10

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)；1-2 µg(ICC)；1-2 µg(IF)；1-3 µg/mL(WB)；0.25 µg/mL(Array)

产品详细信息This antibody is predicted to react with mouse, rat, non-human primate and rabbit 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 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：

Phospho-Histone H3 (Ser10) Antibody (701258) in IFImmunofluorescent analysis of Histone H3 (pS10) was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 15 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with Histone H3 (pS10) Recombinant Rabbit Monoclonal Antibody (Product # 701258) at 1 µg-2 µg 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 Histone H3 (pS10). Panel e shows competition with Histone H3 (pS10) peptide. The images were captured at 20X magnification. Phospho-Histone H3 (Ser10) Antibody (701258) in WBWestern blot analysis of Histone H3 (pS10) was performed by loading 20 µg of HeLa (lane1) and HeLa treated for 25 minutes with 100 microM of Calyculin A (lane2) cell lysates using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Histone H3 (pS10) was detected at ~17 kDa using Histone H3 (pS10) Recombinant Rabbit Monoclonal Antibody (Product # 701258) at 1 µg-3 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. To confirm specificity, competition was performed with phosphopeptide (10 µg/mL) as shown in corresponding blot on right. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

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参考文献：

1. Journal of negative results in biomedicineSubstrate stiffness effect and chromosome missegregation in hIPS cells.

技术参数

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