製品の概要

  • 製品名Anti-PHF8 antibody - ChIP Grade
    PHF8 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to PHF8 - ChIP Grade
  • アプリケーション適用あり: WB, ICC/IF, IHC-P, ChIPmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 800 - 900 of Human PHF8.

    (Peptide available as ab36067.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; U2OS; Jurkat; HepG2.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab36068 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
WB
ICC/IF
IHC-P
ChIP
  • 追加情報ICC/IF: Use at a concentration of 5 µg/ml.
    IHC-P: Use at a concentration of 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
    WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 125 & 140 kDa (predicted molecular weight: 118 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • ターゲット情報

    • 機能Histone lysine demethylase with selectivity for the di-and monomethyl states that plays a key role cell cycle progression, rDNA transcription and brain development. Demethylates mono- and dimethylated histone H3 'Lys-9' residue (H3K9Me1 and H3K9Me2), dimethylated H3 'Lys-27' (H3K27Me2) and monomethylated histone H4 'Lys-20' residue (H4K20Me1). Acts as a transcription activator as H3K9Me1, H3K9Me2, H3K27Me2 and H4K20Me1 are epigenetic repressive marks. Involved in cell cycle progression by being required to control G1-S transition. Acts as a coactivator of rDNA transcription, by activating polymerase I (pol I) mediated transcription of rRNA genes. Required for brain development, probably by regulating expression of neuron-specific genes. Only has activity toward H4K20Me1 when nucleosome is used as a substrate and when not histone octamer is used as substrate. May also have weak activity toward dimethylated H3 'Lys-36' (H3K36Me2), however, the relevance of this result remains unsure in vivo. Specifically binds trimethylated 'Lys-4' of histone H3 (H3K4me3), affecting histone demethylase specificity: has weak activity toward H3K9Me2 in absence of H3K4me3, while it has high activity toward H3K9me2 when binding H3K4me3.
    • 関連疾患Defects in PHF8 are the cause of mental retardation syndromic X-linked Siderius type (MRXSSD) [MIM:300263]. A disorder characterized by mild to borderline mental retardation with or without cleft lip/cleft palate.
    • 配列類似性Belongs to the JHDM1 histone demethylase family. JHDM1D subfamily.
      Contains 1 JmjC domain.
      Contains 1 PHD-type zinc finger.
    • ドメインThe PHD-type zinc finger mediates the binding to H3K4me3. Binding to H3K4me3 promotes its access to H3K9me2.
      The linker region is a critical determinant of demethylase specificity. It enables the active site of JmjC to reach the target H3K9me2 when the PHD-type zinc finger binds to H3K4me3.
    • 翻訳後修飾Phosphorylation at Ser-69 and Ser-120 are required for dissociation from chromatin and accumulation of H4K20Me1 levels during prophase.
    • 細胞内局在Nucleus. Nucleus > nucleolus. Recruited to H3K4me3 sites on chromatin during interphase. Dissociates from chromatin when cells enter mitosis.
    • Information by UniProt
    • 参照データベース
    • 別名
      • Histone lysine demethylase PHF8 antibody
      • JHDM1F antibody
      • Jumonji C domain containing histone demethylase 1F antibody
      • MRXSSD antibody
      • PHD finger protein 8 antibody
      • PHF8 antibody
      • PHF8_HUMAN antibody
      • ZNF422 antibody
      see all

    Anti-PHF8 antibody - ChIP Grade 画像

    • All lanes : Anti-PHF8 antibody - ChIP Grade (ab36068) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
      Lane 4 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 118 kDa
      Observed band size : 140 kDa (why is the actual band size different from the predicted?)


      Exposure time : 2 minutes
    • All lanes : Anti-PHF8 antibody - ChIP Grade (ab36068) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
      Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 118 kDa
      Observed band size : 125,140 kDa (why is the actual band size different from the predicted?)
    • ICC/IF image of ab36068 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab36068, 5µg/ml) 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HepG2 cells at 5µg/ml, and in 100% methanol fixed (5 min) Hek293 cells at 5µg/ml.
    • IHC image of ab36068 staining in human skin 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 ab36068, 5µg/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.

    Anti-PHF8 antibody - ChIP Grade (ab36068) 使用論文

    This product has been referenced in:
    • Castillo J  et al. Genomic and proteomic dissection and characterization of the human sperm chromatin. Mol Hum Reprod 20:1041-53 (2014). Read more (PubMed: 25193639) »
    • Sun X  et al. Oncogenic Features of PHF8 Histone Demethylase in Esophageal Squamous Cell Carcinoma. PLoS One 8:e77353 (2013). WB, IHC-P ; Human . Read more (PubMed: 24146981) »

    See all 5 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (4-20%)
    Sample Human Cell lysate - whole cell (293T)
    Specification 293T
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    投稿 Nov 07 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
    Sample Mouse Cell (Primary cells, testicular germ cells)
    Specification Primary cells, testicular germ cells
    Permeabilization Yes - Triton X-100
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Jul 30 2013

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab84779.

    To check the status of the order please contact o...

    Read More

    Thank you for your reply. I am glad to hear your are able to get the correct band.

    Cross linking should not interfere with the IP; however cross-linking is a time-critical procedure. Cross-linking should generally only be carried out for a f...

    Read More

    Thank you for contacting us. I am sorry to hear that you have been experiencing issues when in ChIP using this product. According to the Uniprot information for this protein (http://www.uniprot.org/uniprot/Q80TJ7)this product may recognize the ~1...

    Read More
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application ChIP
    Sample Human Cell lysate - nuclear (T-cell leukemia (SupT1))
    Specification T-cell leukemia (SupT1)
    Type Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde 1%
    Detection step Real-time PCR
    Positive control Transcriptional Start site (TSS)
    Negative control PHF8 specific shRNA treated cells
    Username

    Abcam user community

    Verified customer

    投稿 Apr 18 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (HPB-ALL (T-cell Leukemia), HeLa cells)
    Loading amount 20 µg
    Specification HPB-ALL (T-cell Leukemia), HeLa cells
    Treatment PHF8 specific shRNA
    Gel Running Conditions Reduced Denaturing (7.5%)
    Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    投稿 Apr 10 2012

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