Key features and details
- Mouse monoclonal [3B1] to CUG-BP1
- Suitable for: IHC-FoFr, IHC-P, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Rabbit, Cow, Human, Pig
- Isotype: IgG1
Product nameAnti-CUG-BP1 antibody [3B1]
See all CUG-BP1 primary antibodies
DescriptionMouse monoclonal [3B1] to CUG-BP1
Tested applicationsSuitable for: IHC-FoFr, IHC-P, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Rat, Rabbit, Cow, Human, Pig
corresponding to CUG-BP1.
- WB: HeLa, HEK-293T and SHSY-5Y cell lysates; Human kidney and brain tissue lysates. IF/ICC: MCF7 cells. IHC: Human Breast cancer tissue
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
PurityProtein G purified
Light chain typekappa
KO cell lysates
Our Abpromise guarantee covers the use of ab9549 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: 1/500. Detects a band of approximately 50 kDa in HeLa lysates(predicted molecular weight: 54 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionRNA-binding protein implicated in the regulation of several post-transcriptional events. Involved in pre-mRNA alternative splicing, mRNA translation and stability. Mediates exon inclusion and/or exclusion in pre-mRNA that are subject to tissue-specific and developmentally regulated alternative splicing. Specifically activates exon 5 inclusion of cardiac isoforms of TNNT2 during heart remodeling at the juvenile to adult transition. Acts as both an activator and repressor of a pair of coregulated exons: promotes inclusion of the smooth muscle (SM) exon but exclusion of the non-muscle (NM) exon in actinin pre-mRNAs. Activates SM exon 5 inclusion by antagonizing the repressive effect of PTB. Promotes exclusion of exon 11 of the INSR pre-mRNA. Inhibits, together with HNRNPH1, insulin receptor (IR) pre-mRNA exon 11 inclusion in myoblast. Increases translation and controls the choice of translation initiation codon of CEBPB mRNA. Increases mRNA translation of CEBPB in aging liver (By similarity). Increases translation of CDKN1A mRNA by antagonizing the repressive effect of CALR3. Mediates rapid cytoplasmic mRNA deadenylation. Recruits the deadenylase PARN to the poly(A) tail of EDEN-containing mRNAs to promote their deadenylation. Required for completion of spermatogenesis (By similarity). Binds to (CUG)n triplet repeats in the 3'-UTR of transcripts such as DMPK and to Bruno response elements (BREs). Binds to muscle-specific splicing enhancer (MSE) intronic sites flanking the alternative exon 5 of TNNT2 pre-mRNA. Binds to AU-rich sequences (AREs or EDEN-like) localized in the 3'-UTR of JUN and FOS mRNAs. Binds to the IR RNA. Binds to the 5'-region of CDKN1A and CEBPB mRNAs. Binds with the 5'-region of CEBPB mRNA in aging liver.
Sequence similaritiesBelongs to the CELF/BRUNOL family.
Contains 3 RRM (RNA recognition motif) domains.
modificationsPhosphorylated. Its phosphorylation status increases in senescent cells.
Cellular localizationNucleus. Cytoplasm. RNA-binding activity is detected in both nuclear and cytoplasmic compartments.
- Information by UniProt
- 50 kDa Nuclear polyadenylated RNA binding protein antibody
- 50 kDa nuclear polyadenylated RNA-binding protein antibody
- Bruno like 2 antibody
All lanes : Anti-CUG-BP1 antibody [3B1] (ab9549) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : CELF1 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Human brain tissue lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 52 kDa why is the actual band size different from the predicted?
Lanes 1-4: Merged signal (red and green). Green - ab9549 observed at 52 kDa. Red - loading control ab181602 observed at 36 kDa.
ab9549 Anti-CUG-BP1 antibody [3B1] was shown to specifically react with CUG-BP1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266086 (knockout cell lysate ab257390) was used. Wild-type and CUG-BP1 knockout samples were subjected to SDS-PAGE. ab9549 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) 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® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
ab9549 stained MCF7 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9549 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
All lanes : Anti-CUG-BP1 antibody [3B1] (ab9549) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 3 : Human kidney tissue lysate - total protein (ab30203)
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: 20 minutes
IHC image of CUG-BP1 staining in human normal kidney formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9549, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab9549 has been referenced in 16 publications.
- Idris M et al. The MBNL/CELF Splicing Factors Regulate Cytosolic Sulfotransferase 4A1 Protein Expression during Cell Differentiation. Drug Metab Dispos 47:314-319 (2019). PubMed: 30606728
- Rong Z et al. Quantitative Studies of Muscleblind Proteins and Their Interaction With TCF4 RNA Foci Support Involvement in the Mechanism of Fuchs' Dystrophy. Invest Ophthalmol Vis Sci 60:3980-3991 (2019). PubMed: 31560764
- Siddam AD et al. The RNA-binding protein Celf1 post-transcriptionally regulates p27Kip1 and Dnase2b to control fiber cell nuclear degradation in lens development. PLoS Genet 14:e1007278 (2018). PubMed: 29565969
- Nutter CA et al. Developmentally regulated alternative splicing is perturbed in type 1 diabetic skeletal muscle. Muscle Nerve 56:744-749 (2017). PubMed: 28164326
- Bai Z et al. Dynamic transcriptome changes during adipose tissue energy expenditure reveal critical roles for long noncoding RNA regulators. PLoS Biol 15:e2002176 (2017). WB . PubMed: 28763438