Anti-CARM1 antibody (ab128851)
Key features and details
- Rabbit polyclonal to CARM1
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-CARM1 antibody
See all CARM1 primary antibodies -
Description
Rabbit polyclonal to CARM1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human
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Immunogen
Synthetic peptide corresponding to Human CARM1 aa 550 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab129212) -
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
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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 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
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
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
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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
modificationsAuto-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. - Information by UniProt
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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
see all -
Alternative names
- carm1 antibody
- CARM1_HUMAN antibody
- Coactivator associated arginine methyltransferase 1 antibody
see all
Images
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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.
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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.
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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 kDaLanes 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.
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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 secondsThis 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
Datasheets and documents
References (2)
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
