Anti-Histone H1x抗体- ChIP Grade (ab31972)

概述

  • 产品名称Anti-Histone H1x抗体- ChIP Grade
    参阅全部 Histone H1x 一抗
  • 描述
    兔多克隆抗体to Histone H1x - ChIP Grade
  • 特异性Using western blot analysis ab31972 recognises histone H1x in HeLa nuclear extract.
  • 经测试应用适用于: ChIP, ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse
  • 免疫原

    Synthetic peptide corresponding to Human Histone H1x aa 200 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: Q92522
    (Peptide available as ab18052)

  • 阳性对照
    • HeLa nuclear extract This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma.

性能

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应用

Our Abpromise guarantee covers the use of ab31972 in the following tested applications.

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

应用 Ab评论 说明
ChIP Use 2 µg for 25 µg of chromatin.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).

靶标

Anti-Histone H1x antibody - ChIP Grade 图像

  • IHC image of Histone H1x staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. 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 ab31972, 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 haematoxylin and mounted with DPX.

    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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab31972 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.

  • All lanes : Anti-Histone H1x antibody - ChIP Grade (ab31972) at 1 µg/ml

    Lane 1 : HeLa nuclear lysate
    Lane 2 : HeLa nuclear lysate with Human Histone H1x peptide (ab18052) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.


    Predicted band size : 35 kDa
    Observed band size : 35 kDa
    Additional bands at : 98 kDa (possible non-specific secondary antibody binding).
    ab31972 recognises histone H1.X in HeLa nuclear extract at 35 kDa (lane 1), which is blocked using the immunizing peptide ab18052.
  • ICC/IF image of ab31972 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31972, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).

Anti-Histone H1x antibody - ChIP Grade (ab31972)参考文献

This product has been referenced in:
  • Millán-Ariño L  et al. Mapping of six somatic linker histone H1 variants in human breast cancer cells uncovers specific features of H1.2. Nucleic Acids Res 42:4474-93 (2014). ChIP ; Human . Read more (PubMed: 24476918) »
  • Terme JM  et al. Histone H1 Variants Are Differentially Expressed and Incorporated into Chromatin during Differentiation and Reprogramming to Pluripotency. J Biol Chem 286:35347-57 (2011). WB ; Human . Read more (PubMed: 21852237) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (u2os)
Gel Running Conditions Reduced Denaturing (4-20%)
Loading amount 20 µg
Specification u2os
Username

Abcam user community

Verified customer

提交于 Sep 29 2016

Application ChIP
Detection step Semiquantitative PCR
Sample Mouse Cell lysate - whole cell (Mouse embryonic fibroblast)
Specification Mouse embryonic fibroblast
Negative control Preimmune IgG
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control Used our homemade Deaf1 antibody and millipore's MeCP2 antibody for pull down.
Username

Abcam user community

Verified customer

提交于 Feb 10 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MRC5VA - Lung fibroblasts)
Loading amount 15 µg
Specification MRC5VA - Lung fibroblasts
Gel Running Conditions Reduced Denaturing (15% gel)
Blocking step Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. Darren Arbon

Verified customer

提交于 Feb 16 2009

Thank you for your enquiry. I am sorry but to date we do not have information as to the reactivity of either ab31972 or ab17729 with other isoforms of histone H1. These antibodies are relatively new additions to our chromatin range. I appreciate th...

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