Rabbit Polyclonal U2AF65 antibody. Suitable for IP, ChIP, WB, ICC/IF, IHC-P and reacts with Human, Zebrafish, Mouse samples. Cited in 22 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS
Liquid
Polyclonal
IP | ChIP | WB | ICC/IF | IHC-P | |
---|---|---|---|---|---|
Human | Tested | Expected | Tested | Tested | Tested |
Mouse | Predicted | Expected | Predicted | Predicted | Predicted |
Cow | Predicted | Predicted | Predicted | Predicted | Predicted |
Xenopus laevis | Predicted | Predicted | Predicted | Predicted | Predicted |
Zebrafish | Expected | Expected | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Zebrafish | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow, Xenopus laevis, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Zebrafish, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow, Xenopus laevis | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Zebrafish | Dilution info 1/250 | Notes - |
Species Human | Dilution info 1/250 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow, Xenopus laevis, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Zebrafish | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow, Xenopus laevis, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Zebrafish | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Cow, Xenopus laevis, Mouse | Dilution info - | Notes - |
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Plays a role in pre-mRNA splicing and 3'-end processing (PubMed:17024186). By recruiting PRPF19 and the PRP19C/Prp19 complex/NTC/Nineteen complex to the RNA polymerase II C-terminal domain (CTD), and thereby pre-mRNA, may couple transcription to splicing (PubMed:21536736). Induces cardiac troponin-T (TNNT2) pre-mRNA exon inclusion in muscle. Regulates the TNNT2 exon 5 inclusion through competition with MBNL1. Binds preferentially to a single-stranded structure within the polypyrimidine tract of TNNT2 intron 4 during spliceosome assembly. Required for the export of mRNA out of the nucleus, even if the mRNA is encoded by an intron-less gene. Represses the splicing of MAPT/Tau exon 10. Positively regulates pre-mRNA 3'-end processing by recruiting the CFIm complex to cleavage and polyadenylation signals (PubMed:17024186).
U2AF65, U2AF2, Splicing factor U2AF 65 kDa subunit, U2 auxiliary factor 65 kDa subunit, U2 snRNP auxiliary factor large subunit, hU2AF(65), hU2AF65
Rabbit Polyclonal U2AF65 antibody. Suitable for IP, ChIP, WB, ICC/IF, IHC-P and reacts with Human, Zebrafish, Mouse samples. Cited in 22 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS
Liquid
Polyclonal
Affinity purification Immunogen
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
U2AF65 also known as U2 small nuclear RNA auxiliary factor 2 (U2AF2) is a splicing factor structured around 65 kilodaltons in mass. It interacts with RNA as part of the spliceosome to facilitate proper pre-mRNA splicing. The protein ensures the precise recognition and binding of the polypyrimidine tract at the 3' splice site. It is expressed in various human tissues playing an important role in RNA processing across different cell types.
The U2AF65 protein contributes to the assembly of the spliceosome a major RNA-protein complex involved in splicing pre-messenger RNA. It works as part of the heterodimeric U2AF complex along with U2AF35 enhancing the precise selection of splice sites. This activity is essential to ensure accurate and efficient splicing which is necessary for proper gene expression and regulation.
The activity of U2AF65 is an integral part of the RNA splicing pathway which plays a critical role in modulating gene expression. U2AF65 acts in coordination with other splicing factors such as SF1 and SF3B1 ensuring the correct assembly of the functional spliceosome. The accurate regulation of this pathway is essential for generating diversity in protein expression and for maintaining cell homeostasis.
Misregulation of U2AF65 activity has implications in diseases like cancer and spinal muscular atrophy. Alterations in the expression or function of U2AF65 can lead to aberrant splicing events which affect gene expression profiles. In cancer U2AF65 interacts with various oncogenes and tumor suppressor proteins potentially contributing to tumour progression. In spinal muscular atrophy alterations in splicing factors including U2AF65 influence the expression of survival motor neuron proteins intricately linking U2AF65 to the pathology of these conditions.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Although the predicted band size is 53kDa based on Swiss-prot data, a band of 65kDa has been previously observed. J Biol Chem. 2004 Nov 26;279(48):49773-9 (PMID: 15377657)
All lanes: Western blot - Anti-U2AF65 antibody (ab37530) at 1/250 dilution
Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2: Western blot - Jurkat whole cell lysate (Jurkat whole cell lysate ab7899) at 10 µg
Lane 3: Western blot - A-431 whole cell lysate (A-431 whole cell lysate ab7909) at 10 µg
Lane 4: Western blot - HEK-293 whole cell lysate (HEK-293 whole cell lysate ab7902) at 10 µg
Lane 5: Hep G2 whole cell lysate (ab7900) at 10 µg
Lane 6: MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Lane 7: SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg
Lane 8: U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg
All lanes: Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 65 kDa
IHC image of ab37530 staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab37530, 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.
U2AF65 was immunoprecipitated using 0.5mg SHSY5Y whole cell extract, 5µg of Rabbit polyclonal to U2AF65 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, SHSY5Y whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab37530.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
Band: 65kDa: U2AF65.
All lanes: Immunoprecipitation - Anti-U2AF65 antibody (ab37530)
Predicted band size: 54 kDa
All lanes: Western blot - Anti-U2AF65 antibody (ab37530) at 1/250 dilution
Lane 1: Marker
Lane 2: Zebrafish brain homogenate at 20 µg
Lane 3: Zebrafish heart homogenate at 10 µg
Lane 4: Zebrafish liver homogenate at 10 µg
Lane 5: Zebrafish skeletal muscle homogenate at 10 µg
Lane 6: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
All lanes: Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 53 kDa
Exposure time: 5min
ab37530 staining U2AF65 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab37530 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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