Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PHF8 antibody (ab36068)

Key features and details

  • Rabbit polyclonal to PHF8
  • Suitable for: WB, ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-PHF8 antibody
    See all PHF8 primary antibodies

  • Description

    Rabbit polyclonal to PHF8

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Pmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse

  • Immunogen

    Synthetic peptide corresponding to Human PHF8 aa 800-900 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab36067)

Properties

Applications

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.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 125, 140 kDa (predicted molecular weight: 118 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    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.

  • Involvement in disease

    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.

  • Sequence similarities

    Belongs to the JHDM1 histone demethylase family. JHDM1D subfamily.
    Contains 1 JmjC domain.
    Contains 1 PHD-type zinc finger.

  • Domain

    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.

  • Post-translational
    modifications

    Phosphorylation at Ser-69 and Ser-120 are required for dissociation from chromatin and accumulation of H4K20Me1 levels during prophase.

  • Cellular localization

    Nucleus. Nucleus > nucleolus. Recruited to H3K4me3 sites on chromatin during interphase. Dissociates from chromatin when cells enter mitosis.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 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

Images

  • Western blot - Anti-PHF8 antibody (ab36068)
    Western blot - Anti-PHF8 antibody (ab36068)
    All lanes : Anti-PHF8 antibody (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
    All lanes : 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
  • Immunocytochemistry/ Immunofluorescence - Anti-PHF8 antibody (ab36068)
    Immunocytochemistry/ Immunofluorescence - Anti-PHF8 antibody (ab36068)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody (ab36068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody (ab36068)
    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.
  • Western blot - Anti-PHF8 antibody (ab36068)
    Western blot - Anti-PHF8 antibody (ab36068)
    All lanes : Anti-PHF8 antibody (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
    All lanes : 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?

References (14)

Publishing research using ab36068? Please let us know so that we can cite the reference in this datasheet.

ab36068 has been referenced in 14 publications.

  • Chen X  et al. Phf8 histone demethylase deficiency causes cognitive impairments through the mTOR pathway. Nat Commun 9:114 (2018). IHC (PFA fixed) ; Mouse . PubMed: 29317619
  • Ying Z  et al. Short-Term Mitochondrial Permeability Transition Pore Opening Modulates Histone Lysine Methylation at the Early Phase of Somatic Cell Reprogramming. Cell Metab 28:935-945.e5 (2018). PubMed: 30174306
  • Shao P  et al. Histone demethylase PHF8 promotes epithelial to mesenchymal transition and breast tumorigenesis. Nucleic Acids Res 45:1687-1702 (2017). ChIP ; Human . PubMed: 27899639
  • Asensio-Juan E  et al. The histone demethylase PHF8 is a molecular safeguard of the IFN? response. Nucleic Acids Res 45:3800-3811 (2017). WB, ChIP ; Human . PubMed: 28100697
  • Walsh RM  et al. Phf8 loss confers resistance to depression-like and anxiety-like behaviors in mice. Nat Commun 8:15142 (2017). WB, ChIP ; Mouse . PubMed: 28485378
View all Publications for this product

Images

  • Western blot - Anti-PHF8 antibody (ab36068)
    Western blot - Anti-PHF8 antibody (ab36068)
    All lanes : Anti-PHF8 antibody (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
    All lanes : 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
  • Immunocytochemistry/ Immunofluorescence - Anti-PHF8 antibody (ab36068)
    Immunocytochemistry/ Immunofluorescence - Anti-PHF8 antibody (ab36068)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody (ab36068)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHF8 antibody (ab36068)
    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.
  • Western blot - Anti-PHF8 antibody (ab36068)
    Western blot - Anti-PHF8 antibody (ab36068)
    All lanes : Anti-PHF8 antibody (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
    All lanes : 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?

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