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Cell Biology Cell Cycle Cell Cycle Inhibitors p53
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Validated using a knockout cell line

Anti-CARM1 antibody (ab128851)

  • Datasheet
Reviews (1) Submit a question References (2)

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Western blot - Anti-CARM1 antibody (ab128851)
  • Immunocytochemistry/ Immunofluorescence - Anti-CARM1 antibody (ab128851)
  • Western blot - Anti-CARM1 antibody (ab128851)
  • Western blot - Anti-CARM1 antibody (ab128851)

Key features and details

  • Rabbit polyclonal to CARM1
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

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Protein
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Recombinant Human CARM1 protein (ab161025)
Primary
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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

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Overview

  • Product name

    Anti-CARM1 antibody
    See all CARM1 primary antibodies
  • Description

    Rabbit polyclonal to CARM1
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human CARM1 aa 550 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab129212)

  • Positive control

    • WB: HT-29, Caco-2, HEK293T, HeLa, MCF7, MDA MB 231, Jurkat, PC3 and DU145 cell lysates; HeLa Histone Nuclear lysaes. ICC/IF: DI145 cells.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • p53
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Co-activators/co-repressors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cell Cycle Inhibitors
    • p53
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Co-factors
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Methylation
    • Arginine Methylation
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • p53 pathway
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Cofactors, Vitamins / minerals
    • Co-factors

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • KO cell lines

    • Human CARM1 knockout HEK-293T cell line (ab266557)
  • KO cell lysates

    • Human CARM1 knockout HEK-293T cell lysate (ab257871)
  • Recombinant Protein

    • Recombinant Human CARM1 protein (ab161025)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab128851 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
WB
Human
All applications
Mouse
Rat
Horse
Cow
Dog
Pig
Macaque monkey
Gorilla
Orangutan
Application Abreviews Notes
WB (1)
Use a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa).
ICC/IF
Use a concentration of 1 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa).
ICC/IF
Use a concentration of 1 µg/ml.

Target

  • Function

    Methylates (mono- and asymmetric dimethylation) the guanidino nitrogens of arginyl residues in several proteins involved in DNA packaging, transcription regulation, pre-mRNA splicing, and mRNA stability. Recruited to promoters upon gene activation together with histone acetyltransferases from EP300/P300 and p160 families, methylates histone H3 at 'Arg-17' (H3R17me), forming mainly asymmetric dimethylarginine (H3R17me2a), leading to activate transcription via chromatin remodeling. During nuclear hormone receptor activation and TCF7L2/TCF4 activation, acts synergically with EP300/P300 and either one of the p160 histone acetyltransferases NCOA1/SRC1, NCOA2/GRIP1 and NCOA3/ACTR or CTNNB1/beta-catenin to activate transcription. During myogenic transcriptional activation, acts together with NCOA3/ACTR as a coactivator for MEF2C. During monocyte inflammatory stimulation, acts together with EP300/P300 as a coactivator for NF-kappa-B. Acts as coactivator for PPARG, promotes adipocyte differentiation and the accumulation of brown fat tissue. Plays a role in the regulation of pre-mRNA alternative splicing by methylation of splicing factors. Also seems to be involved in p53/TP53 transcriptional activation. Methylates EP300/P300, both at 'Arg-2142', which may loosen its interaction with NCOA2/GRIP1, and at 'Arg-580' and 'Arg-604' in the KIX domain, which impairs its interaction with CREB and inhibits CREB-dependent transcriptional activation. Also methylates arginine residues in RNA-binding proteins PABPC1, ELAVL1 and ELAV4, which may affect their mRNA-stabilizing properties and the half-life of their target mRNAs.
  • Tissue specificity

    Overexpressed in prostate adenocarcinomas and high-grade prostatic intraepithelial neoplasia.
  • Sequence similarities

    Belongs to the protein arginine N-methyltransferase family.
  • Post-translational
    modifications

    Auto-methylated on Arg-550. Methylation enhances transcription coactivator activity. Methylation is required for its role in the regulation of pre-mRNA alternative splicing.
    Phosphorylation at Ser-216 interferes with S-adenosyl-L-methionine binding and strongly reduces methyltransferase activity (By similarity). Phosphorylation at Ser-216 is strongly increased during mitosis, and decreases rapidly to a very low, basal level after entry into the G1 phase of the cell cycle. Phosphorylation at Ser-216 may promote location in the cytosol.
  • Cellular localization

    Nucleus. Cytoplasm. Mainly nuclear during the G1, S and G2 phases of the cell cycle. Cytoplasmic during mitosis, after breakup of the nuclear membrane.
  • Target information above from: UniProt accession Q86X55 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10498 Human
    • Entrez Gene: 59035 Mouse
    • Entrez Gene: 363026 Rat
    • Omim: 603934 Human
    • SwissProt: Q86X55 Human
    • SwissProt: Q9WVG6 Mouse
    • SwissProt: Q4AE70 Rat
    • Unigene: 323213 Human
    • Unigene: 178115 Mouse
    see all
  • Alternative names

    • carm1 antibody
    • CARM1_HUMAN antibody
    • Coactivator associated arginine methyltransferase 1 antibody
    • Coactivator-associated arginine methyltransferase 1 antibody
    • Histone arginine methyltransferase CARM 1 antibody
    • Histone arginine methyltransferase CARM1 antibody
    • Histone-arginine methyltransferase CARM1 antibody
    • PRMT 4 antibody
    • PRMT4 antibody
    • Protein arginine methyltransferase antibody
    • Protein arginine N methyltransferase 4 antibody
    • Protein arginine N-methyltransferase 4 antibody
    see all

Images

  • Western blot - Anti-CARM1 antibody (ab128851)
    Western blot - Anti-CARM1 antibody (ab128851)
    All lanes : Anti-CARM1 antibody (ab128851) at 1 µg/ml

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : CARM1 knockout HEK-293T cell lysate
    Lane 3 : HT-29 cell lysate
    Lane 4 : Caco-2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 66 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab128851 observed at 65 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab128851 was shown to react with CARM1 in wild-type HEK-293T cells in western blot with loss of signal observed in CARM1 knockout cell line ab266557 (CARM1 knockout cell lysate ab257871). Wild-type and CARM1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab128851 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-CARM1 antibody (ab128851)
    Immunocytochemistry/ Immunofluorescence - Anti-CARM1 antibody (ab128851)

    ICC/IF image of ab128851 stained DU145 cells. The cells were 4% formaldehyde fixed (10 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 ab128851 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.

  • Western blot - Anti-CARM1 antibody (ab128851)
    Western blot - Anti-CARM1 antibody (ab128851)
    All lanes : Anti-CARM1 antibody (ab128851) at 1/500 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : CARM1 knockout HEK293T cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 66 kDa
    Observed band size: 66 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab128851 observed at 66 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab128851 Anti-CARM1 antibody was shown to specifically react with CARM1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266557 (knockout cell lysate ab257871) was used.  Wild-type and CARM1 knockout samples were subjected to SDS-PAGE.  ab128851 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-CARM1 antibody (ab128851)
    Western blot - Anti-CARM1 antibody (ab128851)
    All lanes : Anti-CARM1 antibody (ab128851) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lane 3 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lane 4 : HeLa Histone Preparation Nuclear Lysate

    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Lane 6 : PC3 (Human prostate carcinoma cell line) Whole Cell Lysate Whole Cell Lysate
    Lane 7 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate


    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 66 kDa


    Exposure time: 30 seconds


    This antibody was raised against an immunogen that is predicted to recognize isoforms 1 and 3 of CARM1. The predicted molecular weights of isoforms 1 and 3 are 63 kDa and 66 kDa respectively.

Protocols

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (2)

    Publishing research using ab128851? Please let us know so that we can cite the reference in this datasheet.

    ab128851 has been referenced in 2 publications.

    • Li S  et al. The Overexpression of CARM1 Promotes Human Osteosarcoma Cell Proliferation through the pGSK3ß/ß-Catenin/cyclinD1 Signaling Pathway. Int J Biol Sci 13:976-984 (2017). PubMed: 28924379
    • Franek M  et al. Nucleolar Reorganization Upon Site-Specific Double-Strand Break Induction. J Histochem Cytochem 64:669-686 (2016). PubMed: 27680669

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-CARM1 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (ES cells)
    Gel Running Conditions
    Reduced Denaturing (10 % gel)
    Loading amount
    10 µg
    Specification
    ES cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Christel Krueger

    Verified customer

    Submitted Aug 12 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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