Anti-Histone H4 抗体 [mAbcam 17036] - ChIP Grade (ab17036)

製品の概要

  • 製品名Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade
    Histone H4 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [mAbcam 17036] to Histone H4 - ChIP Grade
  • アプリケーション適用あり: IHC-P, ChIP, ICC/IF, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Cow, Human
  • 免疫原

    Synthetic peptide corresponding to Human Histone H4 aa 50 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P62805
    (Peptide available as ab13843)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following: Calf thymus histone preparation; HeLa histone preparation; Histone H4 recombinant protein; NIH3T3 Whole Cell Lysate; MEF1 Whole Cell Lysate; PC12 Whole Cell Lysate.

製品の特性

  • 製品の状態Liquid
  • 保存方法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.
  • バッファーpH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Some batches contain 0.4M arginine stabilisng agent
  • Concentration information loading...
  • 精製度IgG fraction
  • ポリ/モノモノクローナル
  • クローン名mAbcam 17036
  • ミエローマSp2/0-Ag14
  • アイソタイプIgG2a
  • 軽鎖の種類kappa
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab17036 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
IHC-P Use a concentration of 1 µg/ml.
ChIP Use 5 µg for 25 µg of chromatin.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).Can be blocked with Human Histone H4 peptide (ab13843).

We recommend using 3% milk as the blocking agent for Western blot.

ターゲット情報

Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade 画像

  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 2 µg/ml

    Lane 1 : Histone H1 Recombinant Protein
    Lane 2 : Histone H2A Recombinant Protein
    Lane 3 : Histone H2B Recombinant Protein
    Lane 4 : Histone H3.1 Recombinant Protein
    Lane 5 : Histone H4 Recombinant Protein
    Lane 6 : Histone H1 Recombinant Protein
    Lane 7 : Histone H2A Recombinant Protein
    Lane 8 : Histone H2B Recombinant Protein
    Lane 9 : Histone H3.1 Recombinant Protein
    Lane 10 : Histone H4 Recombinant Protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 11 kDa


    Exposure time : 4 minutes

    Lanes 1-5: 1% BSA blocking buffer

    Lanes 6-10: 3% Milk blocking buffer

    We recommend using 3% milk as the blocking agent for Western blot.

  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 5 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
    Lane 2 : HeLa Histone Preparation Nuclear Lysate at 2.5 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 1 µg/ml

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • 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, 5µg of ab17036 (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.
  • ICC/IF image of ab17036 stained human HepG2 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab17036, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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). This antibody also gave a positive IF result in HeLa and HEK 293 cells.

  • IHC image of ab17036 staining in Human Hodgekin's Lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab17036, 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.
  • All lanes : Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) at 5 µg/ml

    Lane 1 : Histone H1 Recombinant Protein
    Lane 2 : Histone H2A Recombinant Protein
    Lane 3 : Histone H2B Recombinant Protein
    Lane 4 : Histone H3.1 Recombinant Protein
    Lane 5 : Histone H4 Recombinant Protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 11 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

Anti-Histone H4 antibody [mAbcam 17036] - ChIP Grade (ab17036) 使用論文

This product has been referenced in:
  • Dikopoltsev E  et al. FBXO22 protein is required for optimal synthesis of the N-methyl-D-aspartate (NMDA) receptor coagonist D-serine. J Biol Chem 289:33904-15 (2014). Read more (PubMed: 25336657) »

See 1 Publication for this product

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