Overview

  • Product nameAnti-PRMT5 antibody [EPR5772]
    See all PRMT5 primary antibodies
  • Description
    Rabbit monoclonal [EPR5772] to PRMT5
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PRMT5 aa 50-150.

  • Positive control
    • WB: HEK293, HepG2, HeLa, and NIH3T3 cell lysates; mouse and rat brain tissue lysate. ICC/IF: HepG2 and HeLa cells. IHC-P: human infiltrating duct carcinoma of breast tissue, mouse liver tissue. Flow Cyt: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

Applications

Our Abpromise guarantee covers the use of ab109451 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 1/10000 - 1/50000. Detects a band of approximately 72 kDa (predicted molecular weight: 73 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/100 - 1/250.

Flow Cyt 1/100.

For unpurified use at 1/500 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/50.

For purifed use at 1/50

For unpurified use at 1/100 - 1/250.

  • Application notesIs unsuitable for IP.
  • Target

    • FunctionArginine methyltransferase that can both catalyze the formation of omega-N monomethylarginine (MMA) and symmetrical dimethylarginine (sDMA), with a preference for the formation of MMA. Specifically mediates the symmetrical dimethylation of arginine residues in the small nuclear ribonucleoproteins Sm D1 (SNRPD1) and Sm D3 (SNRPD3); such methylation being required for the assembly and biogenesis of snRNP core particles. Methylates SUPT5H. Mono- and dimethylates arginine residues of myelin basic protein (MBP) in vitro. Plays a role in the assembly of snRNP core particles. May play a role in cytokine-activated transduction pathways. Negatively regulates cyclin E1 promoter activity and cellular proliferation. May regulate the SUPT5H transcriptional elongation properties. May be part of a pathway that is connected to a chloride current, possibly through cytoskeletal rearrangement. Methylates histone H2A and H4 'Arg-3' during germ cell development. Methylates histone H3 'Arg-8', which may repress transcription. Methylates the Piwi proteins (PIWIL1, PIWIL2 and PIWIL4), methylation of Piwi proteins being required for the interaction with Tudor domain-containing proteins and subsequent localization to the meiotic nuage. Methylates RPS10.
    • Tissue specificityUbiquitous.
    • Sequence similaritiesBelongs to the protein arginine N-methyltransferase family.
    • Post-translational
      modifications
      Disulfide bonds and non-covalent association mediate homooligomers formation.
    • Cellular localizationCytoplasm. Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • 72 kDa ICln binding protein antibody
      • 72 kDa ICln-binding protein antibody
      • ANM5_HUMAN antibody
      • Histone synthetic lethal 7, S. cerevisiae, homolog of antibody
      • Histone-arginine N-methyltransferase PRMT5 antibody
      • HMT1 hnRNP methyltransferase like 5 antibody
      • HOMOLOG OF; SKB1 antibody
      • HRMT1L5 antibody
      • IBP72 antibody
      • Jak-binding protein 1 antibody
      • JBP 1 antibody
      • JBP1 antibody
      • PRMT 5 antibody
      • PRMT5 antibody
      • Protein arginine methyltransferase 5 antibody
      • Protein arginine N methyltransferase 5 antibody
      • Protein arginine N methyltransferase 5 N terminally processed antibody
      • Protein arginine N-methyltransferase 5 antibody
      • S. POMBE antibody
      • S. POMBE HOMOLOG OF; SKB1 antibody
      • SHK1 KINASE BINDING PROTEIN 1 antibody
      • Shk1 kinase binding protein 1 homolog antibody
      • Shk1 kinase-binding protein 1 homolog antibody
      • Shk1 kinase/binding protein 1, S. pombe, homolog of antibody
      • SKB 1 antibody
      • SKB1 antibody
      • SKB1 homolog antibody
      • SKB1: SKB1 homolog (S. pombe) antibody
      • SKB1Hs antibody
      see all

    Anti-PRMT5 antibody [EPR5772] images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue sections labelling PRMT5 with purified ab109451 at 1/100. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

    • Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 73 kDa
      Observed band size : 72 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/50000 dilution (purified)

      Lane 1 : HEK293 cell lysate
      Lane 2 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 73 kDa
      Observed band size : 72 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified)

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 73 kDa
      Observed band size : 72 kDa (why is the actual band size different from the predicted?)

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labelling PRMT5 with purified ab109451 at 1/100. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PRMT5 (green) with purified ab109451 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/50) and secondary antibody Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/400).

    • ICC/IF image of ab109451 (unpurified) stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab109451, 1/100 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.

    • Overlay histogram showing HeLa cells stained with ab109451 (unpurified, red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109451, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Equilibrium disassociation constant (KD)

      Click here to learn more about KD

    References for Anti-PRMT5 antibody [EPR5772] (ab109451)

    This product has been referenced in:
    • Gurung B  et al. Menin directly represses gli1 expression independent of canonical hedgehog signaling. Mol Cancer Res 11:1215-22 (2013). Read more (PubMed: 23928057) »
    • Goyal A  et al. Generation of human induced pluripotent stem cells using epigenetic regulators reveals a germ cell-like identity in partially reprogrammed colonies. PLoS One 8:e82838 (2013). ICC/IF ; Human . Read more (PubMed: 24349377) »

    See all 2 Publications for this product

    Product Wall

    Application ChIP
    Sample Mouse Cell lysate - whole cell (Mouse NPCs/NSCs)
    Specification Mouse NPCs/NSCs
    Detection step Real-time PCR
    Type Cross-linking (X-ChIP)
    Duration of cross-linking step: 60 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Bioruptor
    Username

    Mr. Nitin Khandelwal

    Verified customer

    Submitted Jun 19 2015

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (293T, HepG2)
    Loading amount 20 µg
    Specification 293T, HepG2
    Gel Running Conditions Reduced Denaturing (8%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7% · Temperature: 20°C
    Username

    Mr. Dongil Kim

    Verified customer

    Submitted Feb 01 2013

    Thank you for your phone call. I am sorry to hear that these 3 antibodies are not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality....

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"