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RabMAb

Anti-Histone H4 (acetyl K5) 抗体 [EP1000Y] - ChIP Grade (ab51997)

製品の概要

  • 製品名
    Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade
    Histone H4 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP1000Y] to Histone H4 (acetyl K5) - ChIP Grade
  • アプリケーション
    適用あり: ChIP, WB, IHC-P, ICC/IF, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Xenopus laevis, Rice
  • 免疫原

    corresponding to Human Histone H4 aa 1-100 (N terminal) (acetyl K5).
    Database link: P62805

  • ポジティブ・コントロール
    • HeLa, NIH/3T3, C6 cells or human brain glioma, human cervical carcinoma, human normal colon FFPE, mouse liver and rat cerebral cortex tissue.
  • 特記事項

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab51997 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ChIP Use at an assay dependent concentration. PubMed: 18354499
WB 1/500000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).

For unpurified use at 1/10000- 1/50000.

IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/5000.

For unpurified use at 1/250- 1/500.

IP 1/30.

ターゲット情報

画像

  • All lanes : Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997) at 1/500000 dilution (purified)

    Lane 1 : Nuclear extract of HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 7mM Sodium Butyrate for 24 hours
    Lane 2 : Untreated nuclear extract of HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 4 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates
    Lane 5 : C6 (Rat glial tumor cell line) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 6 : Untreated C6 (Rat glial tumor cell line) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 11 kDa
    Observed band size : 11 kDa

    Blocking and diluting buffer: 5% NFDM/TBST.

  • IHC image of unpurified ab51997 staining Histone H4 (acetyl K5) in human colon 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 (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab51997, 1/200 dilution, 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 haematoxylin 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 analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma)treated with 500ng/m Trichostatin A for 4 hours labeling Histone H4 (acetyl K5) with purified ab51997 at 1/5000 dilution (0.1μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab195889, an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 1/200 (2.5 μg/ml). ab150077, a Goat anti rabbit IgG(Alexa Fluor® 488) secondary antibody was used at 1/1000 dilution. PBS instead of the primary antibody was used as a control. DAPI nuclear staining.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

  • All lanes : Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997) at 1/1000000 dilution (unpurified)

    Lane 1 : Untreated HeLa cells
    Lane 2 : TSA treated HeLa calls

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat anti-rabbit HRP labelled (1:2000)

    Predicted band size : 11 kDa
    Observed band size : 11 kDa
  • ab51997 (purified) at 1/30 dilution (2µg) immunoprecipitating Histone H4 (acetyl K5) in HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate.

    Lane 1 (input): HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate 10ug
    Lane 2 (+): ab51997+ HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51997 in HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  • ICC/IF image of unpurtified ab51997 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51997, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

参考文献

This product has been referenced in:
  • Col E  et al. Bromodomain factors of BET family are new essential actors of pericentric heterochromatin transcriptional activation in response to heat shock. Sci Rep 7:5418 (2017). Read more (PubMed: 28710461) »
  • Zhang X  et al. Solar Simulated Ultraviolet Radiation Induces Global Histone Hypoacetylation in Human Keratinocytes. PLoS One 11:e0150175 (2016). WB . Read more (PubMed: 26918332) »

See all 37 Publications for this product

レビューと Q&A

Application
Flow Cytometry
Sample
Mouse Cell (Thymus)
Permeabilization
Yes - FIX & PERM® Cell Fixation & Cell Permeabilization Kit
Gating Strategy
Thymocytes (CD45+ Epcam- cells)
Specification
Thymus
Fixation
FIX & PERM® Cell Fixation & Cell Permeabilization Kit
Username

Abcam user community

Verified customer

投稿 May 20 2015

We listed ChIP on the datasheet based on a reference that used the antibody for this application.

Knutson SK et al. Liver-specific deletion of histone deacetylase 3 disrupts metabolic transcriptional networks. EMBO J 27:1017-28 (2008).
Read More

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Xenopus laevis Cell (Fibroblast)
Specification
Fibroblast
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Apr 05 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Schizosaccharomyces pombe Purified protein (Purified modified histone peptides)
Loading amount
0.1 µg
Specification
Purified modified histone peptides
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

投稿 Feb 03 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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