Recombinant Anti-PML Protein antibody [EPR16792] (ab179466)
Overview
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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 -
Positive control
- WB: 293T, K562, HeLa and A549 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; HAP1. IHC-P: Human mammary gland and breast carcinoma tissues. ICC/IF: K562 cells. IP: K562 whole cell extract.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16792 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lysates
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Recombinant Protein
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Related Products
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 | 1/1000 - 1/2000. Detects a band of approximately 50-110 kDa (predicted molecular weight: 98 kDa). |
Target
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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
modificationsUbiquitinated; 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
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Database links
- Entrez Gene: 5371 Human
- Omim: 102578 Human
- SwissProt: P29590 Human
- Unigene: 526464 Human
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Alternative names
- Acure promyelocytic leukemia, inducer of antibody
- MYL antibody
- Pml antibody
see all
Images
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![Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-384472-anti-pml-protein-antibody-epr16792-western-blot-hela-293t.jpg)
Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PML knockout HeLa cell lysate
Lane 3 : 293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab179466 observed at 50-110 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab179466 Anti-PML Protein antibody [EPR16792] was shown to specifically react with PML Protein in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261811 (knockout cell lysate ab257081) was used. Wild-type and PML Protein knockout samples were subjected to SDS-PAGE. ab179466 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-368594-anti-pml-protein-antibody-epr16792-western-blot-knockout-hap1-pml-hela-hek-293-human.jpg)
Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)All lanes : Anti-PML Protein antibody [EPR16792] (ab179466) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PML knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 98 kDa
Observed band size: 50-110 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab179466 observed at 50-110 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab179466 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in PML knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PML knockout samples were subjected to SDS-PAGE. Ab179466 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-231119-anti-pml-protein-antibody-epr16792-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-231114-anti-pml-protein-antibody-epr16792-western-blot.jpg)
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.
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![Western blot - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-231115-anti-pml-protein-antibody-epr16792-western-blot.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-231116-anti-pml-protein-antibody-epr16792-immunohistochemistry.jpg)
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-231118-anti-pml-protein-antibody-epr16792-immunohistochemistry.jpg)
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunoprecipitation - Anti-PML Protein antibody [EPR16792] (ab179466)](https://www.abcam.com/ps/products/179/ab179466/Images/ab179466-231120-anti-pml-protein-antibody-epr16792-immunoprecipitation.jpg)
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.
Protocols
Datasheets and documents
Certificate of Compliance
References
ab179466 has been referenced in 3 publications.
- Wu W et al. Downregulation of circ_0132266 in chronic lymphocytic leukemia promoted cell viability through miR-337-3p/PML axis. Aging (Albany NY) 11:3561-3573 (2019). PubMed: 31152142
- Komaravelli N et al. Respiratory syncytial virus induces NRF2 degradation through a promyelocytic leukemia protein - ring finger protein 4 dependent pathway. Free Radic Biol Med 113:494-504 (2017). PubMed: 29107745
- Jain A & Vale RD RNA phase transitions in repeat expansion disorders. Nature 546:243-247 (2017). ICC/IF ; Human . PubMed: 28562589
