Recombinant
RabMAb

Recombinant Anti-PRMT5 antibody [EPR5772] (ab109451)

Reviews (2)Q&A (1)References (15)

Overview

  • Product name

    Anti-PRMT5 antibody [EPR5772]
    See all PRMT5 primary antibodies

  • Description

    Rabbit monoclonal [EPR5772] to PRMT5

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human PRMT5 aa 50-150. The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293, HepG2, HeLa, and NIH/3T3 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

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

    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.

    This product is a recombinant rabbit monoclonal antibody.

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 IHC antigen retrieval protocols.

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 notes
    Is unsuitable for IP.

    • Target

    • Function

      Arginine 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 specificity

      Ubiquitous.

    • Sequence similarities

      Belongs to the protein arginine N-methyltransferase family.

    • Post-translational
      modifications

      Disulfide bonds and non-covalent association mediate homooligomers formation.

    • Cellular localization

      Cytoplasm. Nucleus.
    • Information by UniProt

    • Database links

      see all

    • 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

    Images

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

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
      Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : 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/Dilution buffer and concentration: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)Goyal et al PLoS One. 2013 Dec 12;8(12):e82838. doi: 10.1371/journal.pone.0082838. eCollection 2013. Fig 3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Expression and localization of candidate epigenetic reprogramming factors in human embryos.

      Human blastocysts were incubated with primary antibodies for (Panel B) PRMT5 (green), the nuclear DNA stained with DAPI (blue) and visualized by multi-channel confocal microscopy. Note that the expression of PRMT5 primarily localized to the ICM of human blastocysts (indicated by white arrows).

    • Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Anti-PRMT5 antibody [EPR5772] (ab109451) at 1/10000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) 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/Dilution buffer and concentration: 5% NFDM/TBST.

    • Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Western blot - Anti-PRMT5 antibody [EPR5772] (ab109451)
      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
      All lanes : 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/Dilution buffer and concentration: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling 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: Secondary antibody Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/400).

    • Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Immunocytochemistry/ Immunofluorescence - Anti-PRMT5 antibody [EPR5772] (ab109451)

      ICC/IF image of ab109451 (unpurified) stained HepG2 (Human liver hepatocellular carcinoma cell line) cells.

      The cells were fixed with 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize 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 1.43 µM.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] (ab109451)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling 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.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRMT5 antibody [EPR5772] (ab109451)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human infiltrating duct carcinoma of breast tissue sections labeling 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.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Flow Cytometry - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Flow Cytometry - Anti-PRMT5 antibody [EPR5772] (ab109451)

      Overlay histogram showing HeLa (Human liver hepatocellular carcinoma cell line) 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) (1 µg/1x106 cells) used under the same conditions.

      Acquisition of >5,000 events was performed.

    • Other - Anti-PRMT5 antibody [EPR5772] (ab109451)
      Other - Anti-PRMT5 antibody [EPR5772] (ab109451)

      Equilibrium disassociation constant (KD)

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Abe Y  et al. MEP50/PRMT5-mediated methylation activates GLI1 in Hedgehog signalling through inhibition of ubiquitination by the ITCH/NUMB complex. Commun Biol 2:23 (2019). Read more (PubMed: 30675521) »
    • Liu R  et al. PHD finger protein 1 (PHF1) is a novel reader for histone H4R3 symmetric dimethylation and coordinates with PRMT5-WDR77/CRL4B complex to promote tumorigenesis. Nucleic Acids Res 46:6608-6626 (2018). Read more (PubMed: 29846670) »
    See all 15 Publications for this product

    Publishing research using ab109451? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    ChIP abreview for Anti-PRMT5 antibody [EPR5772]

     Good
    Abreviews
    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
    Read More

    Mr. Nitin Khandelwal

    Verified customer

    Submitted Jun 19 2015

    Western blot abreview for Anti-PRMT5 antibody [EPR5772]

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Mr. Dongil Kim

    Verified customer

    Submitted Feb 01 2013

    Question

    samples; HEK293 cells block: 5% milk ab413, ab415: black membrane, protein ladder in white Ab 1/1000 suggestion: use less primary and/or secondary antibody ab109451: lots of bands, Ab 1/1000 suggestion: use less primary antibody (recommended dilution is 1/10,000 to 1/50,000)

    Read More
    Answer

    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. As we discussed over the phone, I would like to offer some suggestions to help optimize the results: For ab413, ab415 I'd suggest using less primary and/or secondary antibody as the membrane shouldn't turn black. This might be an indication of too much antibody on the membrane. For ab109451, most likely using less primary antibody could help with the background as the recommended dilution for the antibody is 1/10,000 to 1/50,000. Should the suggestions not improve the results, please do let me know. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, and I thank you for your cooperation.

    Read More

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