Overview

  • Product nameAnti-PML Protein antibodySee all PML Protein primary antibodies ...
  • Description
    Rabbit polyclonal to PML Protein
  • SpecificityThis antibody detects endogenous levels of total PML protein.
  • Tested applicationsWB, ELISA, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human, African Green Monkey
  • Immunogen

    Synthetic peptide corresponding to Human PML Protein aa 39-53 (N terminal).
    Sequence:

    PSPTERAPASEEEFQ


    Database link: P29590

  • Positive control
    • Extracts from A549 cells Hela cells

Properties

Applications

Our Abpromise guarantee covers the use of ab53773 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 1/500 - 1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 69 kDa).
ELISA 1/10000.
IHC-P Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
IP Use at an assay dependent concentration. PubMed: 23222847

Target

  • FunctionKey component of PML nuclear bodies that regulate a large number of cellular processes by facilitating post-translational modification of target proteins, promoting protein-protein contacts, or by sequestering proteins. Functions as tumor suppressor. Required for normal, caspase-dependent apoptosis in response to DNA damage, FAS, TNF, or interferons. Plays a role in transcription regulation, DNA damage response, DNA repair and chromatin organization. Plays a role in processes regulated by retinoic acid, regulation of cell division, terminal differentiation of myeloid precursor cells and differentiation of neural progenitor cells. Required for normal immunity to microbial infections. Plays a role in antiviral response. In the cytoplasm, plays a role in TGFB1-dependent processes. Regulates p53/TP53 levels by inhibiting its ubiquitination and proteasomal degradation. Regulates activation of p53/TP53 via phosphorylation at 'Ser-20'. Sequesters MDM2 in the nucleolus after DNA damage, and thereby inhibits ubiquitination and degradation of p53/TP53. Regulates translation of HIF1A by sequestering MTOR, and thereby plays a role in neoangiogenesis and tumor vascularization. Regulates RB1 phosphorylation and activity. Required for normal development of the brain cortex during embryogenesis. Can sequester herpes virus and varicella virus proteins inside PML bodies, and thereby inhibit the formation of infectious viral particles. Regulates phosphorylation of ITPR3 and plays a role in the regulation of calcium homeostasis at the endoplasmic reticulum (By similarity). Regulates transcription activity of ELF4. Inhibits specifically the activity of the tetrameric form of PKM2. Together with SATB1, involved in local chromatin-loop remodeling and gene expression regulation at the MHC-I locus. Regulates PTEN compartmentalization through the inhibition of USP7-mediated deubiquitinylation.
  • Involvement in diseaseNote=A chromosomal aberration involving PML may be a cause of acute promyelocytic leukemia (APL). Translocation t(15;17)(q21;q21) with RARA. The PML breakpoints (type A and type B) lie on either side of an alternatively spliced exon.
  • Sequence similaritiesContains 2 B box-type zinc fingers.
    Contains 1 RING-type zinc finger.
  • DomainInteracts with PKM2 via its coiled-coil domain.
    Binds arsenic via the RING-type zinc finger.
  • Post-translational
    modifications
    Ubiquitinated; mediated by RNF4, SIAH1 or SIAH2 and leading to subsequent proteasomal degradation. 'Lys-6'-, 'Lys-11'-, 'Lys-48'- and 'Lys-63'-linked polyubiquitination by RNF4 is polysumoylation-dependent.
    Undergoes 'Lys-11'-linked sumoylation. Sumoylation on all three sites is required for nuclear body formation. Sumoylation on Lys-160 is a prerequisite for sumoylation on Lys-65. The PML-RARA fusion protein requires the coiled-coil domain for sumoylation. Desumoylated by SENP2 and SENP6. Arsenic induces PML and PML-RARA oncogenic fusion proteins polysumoylation and their subsequent RNF4-dependent ubiquitination and proteasomal degradation, and is used as treatment in acute promyelocytic leukemia (APL).
    Phosphorylated in response to DNA damage, probably by ATR.
    Acetylation may promote sumoylation and enhance induction of apoptosis.
  • Cellular localizationNucleus > nucleoplasm. Cytoplasm. Nucleus > PML body. Nucleus > nucleolus. Endoplasmic reticulum membrane. Early endosome membrane. Sumoylated forms localize to the PML nuclear bodies. The B1 box and the RING finger are also required for this nuclear localization. Isoforms lacking a nuclear localization signal are cytoplasmic. Detected in the nucleolus after DNA damage. Sequestered in the cytoplasm by interaction with rabies virus phosphoprotein.
  • Information by UniProt
  • Database links
  • Alternative names
    • Acure promyelocytic leukemia, inducer of antibody
    • MYL antibody
    • PML antibody
    • PML_HUMAN antibody
    • PP8675 antibody
    • Probable transcription factor PML antibody
    • Promyelocytic leukemia antibody
    • Promyelocytic leukemia inducer of antibody
    • Promyelocytic leukemia protein antibody
    • Protein PML antibody
    • RING finger protein 71 antibody
    • RNF 71 antibody
    • RNF71 antibody
    • TRIM 19 antibody
    • TRIM19 antibody
    • Tripartite motif containing protein 19 antibody
    • Tripartite motif protein TRIM19 antibody
    • Tripartite motif-containing protein 19 antibody
    see all

Anti-PML Protein antibody images

  • ab53773 (1/200) staining PML protein in Hela cells (green). Cells were fixed with methanol, permeabilised with triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • Chromatin was prepared from modified murine D3 ES cells (an in vitro model of RB1 imprinted expression was generated by integrating human PPP1R26P1 into intron 2 of mouse Rb1 in murine embryonic stem cells (mESCs)). Cells were fixed with formaldehyde for 10 mins. Chromatin was sheared into fragment sizes ranging from 200 to 600 bp. For immunoprecipitation 50 μl of sheared chromatin was set aside as input and 20-40 μg of sheared chromatin in a volume of 100 μl was used for antibody incubation. 4 µg of ab53773 to PML was used as a non-related control (black bars) and 4 µl of a rabbit polyclonal to RNAPII CTD repeat YSPTSPS (phospho S5) (shaded bars) was added as the test. The immunoprecipitated DNA was quantified at 3 positions on the the retinoblastoma (Rb1) promoter. 1,2 = Rb1_48; 3,4 = Rb1_69 and 5,6 = Rb1_74

  • All lanes : Anti-PML Protein antibody (ab53773) at 1/500 dilution

    Lane 1 : Extracts from A549 cells
    Lane 2 : Extracts from A549 cells, plus immunising peptide


    Predicted band size : 69 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)
  • Ab53773 staining Human normal colon. Staining is localized to the nucleus.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

References for Anti-PML Protein antibody (ab53773)

This product has been referenced in:
  • Bronnimann MP  et al. A transmembrane domain and GxxxG motifs within L2 are essential for papillomavirus infection. J Virol 87:464-73 (2013). Read more (PubMed: 23097431) »
  • Delbarre E  et al. DAXX-dependent supply of soluble (H3.3-H4) dimers to PML bodies pending deposition into chromatin. Genome Res 23:440-51 (2013). IP, ICC/IF . Read more (PubMed: 23222847) »

See all 9 Publications for this product

Product Wall

Application Western blot
Loading amount 15 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES running buffer)
Sample Mouse Tissue lysate - whole (Mouse whole brain tissue lysate, striatum, cortex,)
Specification Mouse whole brain tissue lysate, striatum, cortex,
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Sep 06 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Heat-inactivated normal donkey serum in 0.05% PBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step None
Sample Mouse Tissue sections (Mouse, brain tissue, whole brain sections, striatu)
Specification Mouse, brain tissue, whole brain sections, striatu
Permeabilization Yes - Tween-20
Fixative Paraformaldehyde
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Submitted Jul 22 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Sample African Green Monkey Cell (Cos7 Monkey Kidney Fibroblast)
Specification Cos7 Monkey Kidney Fibroblast
Permeabilization Yes - .3% Triton X100
Fixative Paraformaldehyde
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Submitted Jul 10 2013

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Ovary)
Specification Ovary
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
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Submitted Jul 19 2012

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Testis)
Specification Testis
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C
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Submitted Jul 19 2012

J'ai compris que vous aviez rencontré des difficultés avec deux anti-PML et j'aimerais donc vous rappeler que nos produits sont garantis pour fonctionner tels qu'ils sont décrits sur la fiche technique et que nous pouvons donc touj...

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