Overview

  • Product name
    Anti-PML Protein antibody - N-terminal
    See all PML Protein primary antibodies
  • Description
    Rabbit polyclonal to PML Protein - N-terminal
  • Host species
    Rabbit
  • Specificity
    This antibody detects endogenous levels of total PML protein.
  • Tested applications
    Suitable for: WB, ELISA, IHC-P, ICC/IF, ChIP, 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
    • WB: A549 cell extracts. ICC/IF: HeLa cells. ChIP: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)). IHC-P: Human normal colon tissue.

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.
ChIP Use at an assay dependent concentration. PubMed: 24019952
IP Use at an assay dependent concentration. PubMed: 23222847

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

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

    Lane 1 : A549 (Human lung carcinoma cell line) cell extracts
    Lane 2 : A549 (Human lung carcinoma cell line) cell extracts with immunizing peptide

    Predicted band size: 69 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?

  • ab53773 (1/200) staining PML protein in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (green). Cells were fixed with methanol, permeabilized 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 minutes. 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

  • ab53773 staining PML in human normal colon tissue. Staining is localized to the nucleus.
    Left panel: ab53773 at 4 µg/ml. Right panel: Isotype control.
    Sections were stained using an automated system at room temperature. Sections were rehydrated and antigen retrieved with the EDTA pH 9.0 buffer. Slides were blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with hematoxylin and coverslipped. 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

This product has been referenced in:
  • Diplas BH  et al. The genomic landscape of TERT promoter wildtype-IDH wildtype glioblastoma. Nat Commun 9:2087 (2018). Read more (PubMed: 29802247) »
  • Zink LM  et al. H3.Y discriminates between HIRA and DAXX chaperone complexes and reveals unexpected insights into human DAXX-H3.3-H4 binding and deposition requirements. Nucleic Acids Res 45:5691-5706 (2017). Read more (PubMed: 28334823) »
See all 22 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Huh7 liver)
Specification
Huh7 liver
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 17 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Sep 06 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Jul 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

Submitted Jul 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Jul 19 2012

Answer

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 toujours résoudre un problèmeen vous remboursant, vous faire un avoir, ou vous envoyer un produit de remplacement.

Pourriez-vous, s'il vous plait, me communiquer les informations suivantes :
- la référence de l'anticorps
- le numéro de lot
- le numéro et la date de la commande
- l'application testée
- le type d'échantillon (espèce, lignée cellulaire, tissu)
- quelques détails sur votre protocole (concentration/dilution de l'anticorps, temps d'incubation, agent bloquant, préparation des échantillons, etc.)

Je serai ravi de vous apporter une solution rapide à votre problème.



Autre point, le laboratoire qui produit l'anticorps anti-PML ab67761 m'a informé que malheureusement aucune image Western Blot n'est actuellement disponible. Si vous souhaitezutiliser ab67761 en Western Blot, il me sera possible de vous offrir une offre promotionnelle. Après achat de l'anticorps et l'envoi d’une Abreview avec vos résultats, vous pourrez commander gratuitement n'importe quel anticorps primaire de notre catalogue.
Etapes à suivre :
1. Nous confirmer que vous souhaiteriezutiliser ab67761en Western Blotafin de recevoir le code de remise correspondant. Important, ce code doit être édité avant l’achat de ab67761.
2.Commander ab67761par téléphone, fax ou internet (https://www.abcam.com). Inutile de mentionner le code sur cette commande.
3. Tester l'anticorps
4. Nous faire part de vos résultats grâce à notre système Abreview. Pour plus d’informations https://www.abcam.com/abreviews
5. Après soumission de votre Abreview, appeler notre service clientèle afin de passer votre prochaine commande grâce au code de réduction. Ce code de réduction est utilisable 120 jours après son édition et utilisable lors de l’achat d’un autre anticorps primaire.

N’hésitez pas à nous demander plus d’informations concernant cette offre promotionnelle.

Termes et Conditions : https://www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you for confirming this information and for your help and cooperation with this case.

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

Credit ID: #####

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

Read More

Answer

Merci de nous avoir contactés et désolé pour le délai de ma réponse, j'attendais après le laboratoire les réponses à vos questions.

Concernant l'immunogène : l'information sur la fiche technique est incorrecte. Le peptide utilisé correspond à la régioninterne (entre les acides aminés 400et 500) et non àla région N-terminale. Nous sommes désolés pour cette erreur, la fiche technique va être mise à jour avec la bonneinformation.

J'attends malheureusementtoujours l'image Western Blot pour cet anticorps.

Nous avons2 autresanti-PML garantis pour fonctionner en Western Blot et chez la souris :

ab53773, https://www.abcam.com/ab53773
ab50637, https://www.abcam.com/ab50637



J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

Read More

Answer

Thank you for your message and for providing this further information.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange afree of charge replacementor credit note in compensation.

I look forward to hearing from you with details of how you would like to proceed.

Read More

1-10 of 19 Abreviews or Q&A

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