Recombinant
RabMAb

Anti-PML Protein antibody [EPR16792] (ab179466)

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Overview

  • Product name
    Anti-PML Protein antibody [EPR16792]
    See all PML Protein primary antibodies
  • Description
    Rabbit monoclonal [EPR16792] to PML Protein
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human PML Protein aa 150-400. The exact sequence is proprietary.
    Database link: P29590

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • WB: 293T, K562, HeLa and A549 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates. IHC-P: Human mammary gland and breast carcinoma tissues. ICC/IF: K562 cells. IP: K562 whole cell extract.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab179466 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
IP 1/70.
ICC/IF 1/500.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 50-110 kDa (predicted molecular weight: 98 kDa).

Target

  • Function
    Key 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 disease
    Note=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 similarities
    Contains 2 B box-type zinc fingers.
    Contains 1 RING-type zinc finger.
  • Domain
    Interacts 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 localization
    Nucleus > 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
      • Tripartite motif protein TRIM19 antibody
      • Tripartite motif-containing protein 19 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-PML Protein antibody [EPR16792] (ab179466)
      Immunocytochemistry/ Immunofluorescence - Anti-PML Protein antibody [EPR16792] (ab179466)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling PML Protein with ab179466 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on K562 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

      The negative controls are as follows:-
      -ve control 1 -  ab179466 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
      -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

    • Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)
      Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)
      All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/20000 dilution

      Lane 1 : 293T (Human epithelial cells from embryonic kidney) whole cell lysate
      Lane 2 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
      Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
      Lane 4 : A549 (Human lung carcinoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 98 kDa
      Observed band size: 50-110 kDa (why is the actual band size different from the predicted?)



      Blocking/Dilution buffer: 5% NFDM/TBST.

      ab179466 recognizes 12 isoforms shown as multiple bands ranging from 50 kDa to 110 kDa.

    • Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)
      Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)
      All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/2000 dilution

      Lane 1 : Human fetal brain lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 98 kDa
      Observed band size: 50-110 kDa (why is the actual band size different from the predicted?)



      Blocking/Dilution buffer: 5% NFDM/TBST.

      ab179466 recognizes 12 isoforms shown as multiple bands ranging from 50 kDa to 110 kDa.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] (ab179466)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] (ab179466)

      Immunohistochemical analysis of paraffin-embedded Human mammary gland tissue labeling PML Protein with ab179466 at 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on epithelial cells of Human mammary gland tissue is observed. Counter stained with Hematoxylin.

      Negative control: Used PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] (ab179466)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] (ab179466)

      Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling PML Protein with ab179466 at 1/2000 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stained with Hematoxylin. The breast cancer cells lost expression.

      Reference: Gurrieri C et al. Loss of the Tumor Suppressor PML in Human Cancers of Multiple Histologic Origins. J Natl Cancer Inst 96:269 –279 (2004).

      Negative control: Used PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    • Immunoprecipitation - Anti-PML Protein antibody [EPR16792] (ab179466)
      Immunoprecipitation - Anti-PML Protein antibody [EPR16792] (ab179466)

      PML Protein was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell extract with ab179466 at 1/70 diltuion. Western blot was performed from the immunoprecipitate using ab179466 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: K562 whole cell extract. Lane 2: PBS instead of K562 whole cell extract.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      ab179466 recognizes 12 isoforms shown as multiple bands ranging from 50 kDa to 110 kDa.

    References

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    Customer reviews and Q&As

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    1 Review or Q&A

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    Immunocytochemistry/ Immunofluorescence abreview for Anti-PML Protein antibody [EPR16792]

     Excellent
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U-2OS)
    Permeabilization
    No
    Specification
    U-2OS
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Methanol
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    Submitted Jun 14 2017

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