Anti-Histone H3 (phospho S10) antibody [mAbcam 14955]

• 产品名称

  Anti-Histone H3 (phospho S10)抗体[mAbcam 14955]
  参阅全部 Histone H3 一抗

• 描述

  小鼠单克隆抗体[mAbcam 14955] to Histone H3 (phospho S10)

• 宿主

  Mouse

• 特异性

  ab14955 recognises phospho S10 on Histone H3. It recognises a phospho S28 peptide by ELISA, but is not blocked by phospho S28 in WB.

• 经测试应用

  适用于: IHC-P, ELISA, Flow Cyt (Intra), ICC/IF, WBmore details

• 种属反应性

  与反应: Mouse, Human, Drosophila melanogaster
  预测可用于: Rat, Chicken, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Indian muntjac, Monkey, Schizosaccharomyces pombe, Zebrafish, Mammals, Tobacco, Chlamydomonas reinhardtii, African green monkey, Aspergillus nidulans, Neurospora crassa, Oncopeltus

• 免疫原

  Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

• 阳性对照

• WB: Control HeLa Histone Prep. Colcemid treated HeLa Histone prep. HeLa cell lysate treated with calyculin A. IHC-P: Human kidney tissue. ICC: HeLa and 10T1/2 cells. Flow Cyt (Intra): HeLa cells.

• 常规说明

  Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising tri methyl K9 and phospho S10 dimodified peptide. Clones were also positively screened against both tri methyl K9 and phospho S10 peptides. Clones were negatively screened against the unmodified version of the peptides. This clone binds to the tri methyl K9 and phospho S10 dimodified peptide and to the phospho S10 peptide, but not to the tri methyl K9 peptide or to equivalent unmodified Histone H3 peptide.

  
  

  This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.50
  Preservative: 0.02% Sodium azide
  Constituent: PBS

• 浓度

• 100 µg 浓度为 1 mg/ml

• 纯度

  IgG fraction

• 克隆

  单克隆

• 克隆编号

  mAbcam 14955

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab14955于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

• 数据库链接

  hide

• Entrez Gene: 8358 Human

• Entrez Gene: 8968 Human

• Entrez Gene: 319152 Mouse

• Entrez Gene: 319153 Mouse

• Entrez Gene: 360198 Mouse

• Entrez Gene: 97908 Mouse

• Entrez Gene: 100364501 Rat

• Entrez Gene: 100365669 Rat

• Entrez Gene: 291159 Rat

• Entrez Gene: 679994 Rat

• Entrez Gene: 680511 Rat

• Entrez Gene: 682330 Rat

• Omim: 602810 Human

• SwissProt: P84229 Chicken

• SwissProt: P68431 Human

• SwissProt: P68433 Mouse

• SwissProt: Q6LED0 Rat

• Unigene: 132854 Human

• Unigene: 247813 Human

• Unigene: 247814 Human

• Unigene: 248176 Human

• Unigene: 443021 Human

• Unigene: 484990 Human

• Unigene: 532144 Human

• Unigene: 533292 Human

• Unigene: 546315 Human

• Unigene: 586261 Human

• Unigene: 591778 Human

• Unigene: 221301 Mouse

• Unigene: 261657 Mouse

• Unigene: 377874 Mouse

• Unigene: 390558 Mouse

• Unigene: 397328 Mouse

• Unigene: 138090 Rat

• Entrez Gene: 8350 Human

• Entrez Gene: 8351 Human

• Entrez Gene: 8352 Human

• Entrez Gene: 8353 Human

• Entrez Gene: 8354 Human

• Entrez Gene: 8355 Human

• Entrez Gene: 8356 Human

• Entrez Gene: 8357 Human

• 别名

• H31_HUMAN antibody

• H3F3 antibody

• H3FA antibody

• Hist1h3a antibody

• HIST1H3B antibody

• HIST1H3C antibody

• HIST1H3D antibody

• HIST1H3E antibody

• HIST1H3F antibody

• HIST1H3G antibody

• HIST1H3H antibody

• HIST1H3I antibody

• HIST1H3J antibody

• HIST3H3 antibody

• histone 1, H3a antibody

• Histone cluster 1, H3a antibody

• Histone H3 3 pseudogene antibody

• Histone H3.1 antibody

• Histone H3/a antibody

• Histone H3/b antibody

• Histone H3/c antibody

• Histone H3/d antibody

• Histone H3/f antibody

• Histone H3/h antibody

• Histone H3/i antibody

• Histone H3/j antibody

• Histone H3/k antibody

• Histone H3/l antibody

• H3 histone family member E pseudogene antibody

• H3 histone family, member A antibody

• H3/A antibody

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS only (ab238673).

  ab238673 staining Histone H3 (phospho S10) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab238673 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  IHC image of ab14955 staining Histone H3 (phospho S10) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond.

  The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14955, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  ab14955 staining Histone H3 (phospho S10) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab14955 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 1 µg/ml
  
  Lane 1 : Control HeLa Histone Prep
  Lane 2 : Colcemid treated HeLa Histone prep
  Lane 3 : Control HeLa Histone Prep with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
  Lane 4 : Colcemid treated HeLa Histone prep with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
  Lane 5 : Control HeLa Histone Prep with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
  Lane 6 : Colcemid treated HeLa Histone prep with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
  Lane 7 : Control HeLa Histone Prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
  Lane 8 : Colcemid treated HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
  Lane 9 : Control HeLa Histone Prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
  Lane 10 : Colcemid treated HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
  
  Lysates/proteins at 0.5 µg per lane.
  
  Secondary
  All lanes : Rabbit polyclonal to Mouse IgG H&L (HRP) at 1/5000 dilution
  
  Performed under reducing conditions.
  
  Predicted band size: 15 kDa
  Observed band size: 17 kDawhy is the actual band size different from the predicted?
  
  

• 

  Flow Cytometry (Intracellular) - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)This image is courtesy of an Abreview submitted by Dr Kirk McManus

  ab14955 (1/1000) stainning a population of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells positive for Histone H3 (phospho S10). Cells were trypsizined, pelleted and fixed in ice cold ethanol. Cellular debris was eliminated and the FL2-A/FL2-W was used to eliminate clumping cells. For further experimental details please refer to abreview.

  See Abreview

• 

  Flow Cytometry (Intracellular) - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  Flow cytometry dot plot showing HeLa cells stained with ab14955 (red line). The cells were fixed with 100% ethanol. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab14955) (1x 106 in 100μl at 1μg/ml (1/500)) for 30 min at 22°C. The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/4000 for 30 min at 22°C. DNA staining was performed by pre-incubation with RNase A (250μg/ml) followed by DAPI.

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• 

  ELISA - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  By ELISA, ab14955 detects:
  the singly modified phospho S10 peptide and the dual modified phospho S10 and tri methyl K9 peptide (the orange and grey lines respectively),
  less strongly detects the phospho S28 peptide (purple line),
  does not detect the equivalent non-modified Histone H3 peptide for S10 or the singly modified tri methyl K9 peptide (the 2 lines at the bottom of the figure).
  
  The antibody recognises phospho S28 by ELISA (although a phospho S28 peptide does not block the antibody in Western blotting).

• 

  Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 1/2000 dilution
  
  Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
  Lane 2 : HeLa cell lysate treated with calyculin A
  Lane 3 : HeLa cell lysate treated with calyculin A and alkaline phosphatase
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  All lanes : HRP goat anti-rabbit (H+L) at 1/20000 dilution
  
  Predicted band size: 15 kDa
  
  
  Exposure time: 1 second
  

• 

  Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)

  All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 0.5 µg/ml
  
  Lane 1 : Control HeLa Histone prep
  Lane 2 : Colecemid treated HeLa Histone prep
  Lane 3 : Control HeLa Histone prep with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
  Lane 4 : Colecemid treated HeLa Histone prep with Human Histone H3 (unmodified) peptide (ab7228) at 1 µg/ml
  Lane 5 : Control HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
  Lane 6 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
  Lane 7 : Control HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
  Lane 8 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
  
  Lysates/proteins at 5 µg per lane.
  
  Secondary
  All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
  
  Predicted band size: 15 kDa
  Observed band size: 20 kDawhy is the actual band size different from the predicted?
  
  

• 

  Western blot - Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955)Image courtesy of Richelle Sopko, Harvard University, U.S.A

  All lanes : Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] (ab14955) at 1/1000 dilution
  
  Lane 1 : Wild type 0-4 hour old fruit fly embryo lysate
  Lane 2 : 0-4 hour old fruit fly embryo lysate expressing wee RNAi
  
  Secondary
  All lanes : Anti-mouse IgG, peroxidase-linked at 1/10000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Predicted band size: 15 kDa
  
  
  Exposure time: 5 seconds
  
  
  

  Blocking: 10% BSA

  wee shRNA embryos (lane 2) should display elevated phospho H3Ser10 levels relative to wild type

  See Abreview