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Rabbit Polyclonal U2AF65 antibody. Suitable for IP, ChIP, WB, ICC/IF, IHC-P and reacts with Human, Zebrafish, Mouse samples. Cited in 22 publications.


Images

Western blot - Anti-U2AF65 antibody (AB37530), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-U2AF65 antibody (AB37530), expandable thumbnail
  • Immunoprecipitation - Anti-U2AF65 antibody (AB37530), expandable thumbnail
  • Western blot - Anti-U2AF65 antibody (AB37530), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-U2AF65 antibody (AB37530), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS

Form

Liquid

Clonality

Polyclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Consider this alternative

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPChIPWBICC/IFIHC-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

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Zebrafish

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow, Xenopus laevis, Mouse

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Zebrafish, Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow, Xenopus laevis

Dilution info

-

Notes

-

Tested
Tested

Species

Zebrafish

Dilution info

1/250

Notes

-

Species

Human

Dilution info

1/250

Notes

-

Predicted
Predicted

Species

Cow, Xenopus laevis, Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1 µg/mL

Notes

-

Expected
Expected

Species

Zebrafish

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow, Xenopus laevis, Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

5 µg/mL

Notes

-

Expected
Expected

Species

Zebrafish

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Cow, Xenopus laevis, Mouse

Dilution info

-

Notes

-

Associated Products

Select an associated product type

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Target data

Function

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).

Alternative names

Recommended products

Rabbit Polyclonal U2AF65 antibody. Suitable for IP, ChIP, WB, ICC/IF, IHC-P and reacts with Human, Zebrafish, Mouse samples. Cited in 22 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Polyclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Purification technique

Affinity purification Immunogen

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

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5 product images

  • Western blot - Anti-U2AF65 antibody (ab37530), expandable thumbnail

    Western blot - Anti-U2AF65 antibody (ab37530)

    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

    Secondary

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-U2AF65 antibody (ab37530), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-U2AF65 antibody (ab37530)

    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.

  • Immunoprecipitation - Anti-U2AF65 antibody (ab37530), expandable thumbnail

    Immunoprecipitation - Anti-U2AF65 antibody (ab37530)

    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

  • Western blot - Anti-U2AF65 antibody (ab37530), expandable thumbnail

    Western blot - Anti-U2AF65 antibody (ab37530)

    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

    Secondary

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-U2AF65 antibody (ab37530), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-U2AF65 antibody (ab37530)

    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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