Anti-U2AF65 antibody
| Name | Anti-U2AF65 antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab37530 |
| Prices | $400.00 |
| Sizes | 200 µl |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | IHC-P ICC/IF ICC/IF IP WB |
| Species Reactivities | Mouse, Human, Zebrafish, Bovine, Xenopus |
| Antigen | Synthetic peptide derived from within residues 200 - 300 of Human U2AF65 |
| Description | Rabbit Polyclonal |
| Gene | U2AF2 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilutionLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate (ab7900)Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell LysateLane 8 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryRabbit IgG secondary antibody (ab28446) at 1/10000 dilutionPerformed under reducing conditions.
ICC/IF image of ab37530 stained human HeLa cells. The cells were PFA fixed (10 min) and incubated with the antibody (ab37530, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence. 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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.
All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilutionLane 1 : MarkerLane 2 : Zebrafish brain homogenate at 20 µgLane 3 : Zebrafish heart homogenate at 10 µgLane 4 : Zebrafish liver homogenate at 10 µgLane 5 : Zebrafish skeletal muscle homogenate at 10 µgLane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µgSecondaryGoat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
![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) (ab99697).Band: 65kDa: U2AF65.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/16943_U2AF65-Primary-antibodies-ab37530-5.jpg)
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) (ab99697).Band: 65kDa: U2AF65.
Product References
CTD serine-2 plays a critical role in splicing and termination factor recruitment - CTD serine-2 plays a critical role in splicing and termination factor recruitment
Gu B, Eick D, Bensaude O. Nucleic Acids Res. 2013 Feb 1;41(3):1591-603.
Phosphorylation of the alternative mRNA splicing factor 45 (SPF45) by Clk1 - Phosphorylation of the alternative mRNA splicing factor 45 (SPF45) by Clk1
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A positive feedback loop links circadian clock factor CLOCK-BMAL1 to the basic - A positive feedback loop links circadian clock factor CLOCK-BMAL1 to the basic
Lande-Diner L, Boyault C, Kim JY, Weitz CJ. Proc Natl Acad Sci U S A. 2013 Oct 1;110(40):16021-6. doi:
Perturbation of U2AF65/NXF1-mediated RNA nuclear export enhances RNA toxicity in - Perturbation of U2AF65/NXF1-mediated RNA nuclear export enhances RNA toxicity in
Tsoi H, Lau CK, Lau KF, Chan HY. Hum Mol Genet. 2011 Oct 1;20(19):3787-97.
Splicing-independent recruitment of U1 snRNP to a transcription unit in living - Splicing-independent recruitment of U1 snRNP to a transcription unit in living
Spiluttini B, Gu B, Belagal P, Smirnova AS, Nguyen VT, Hebert C, Schmidt U, Bertrand E, Darzacq X, Bensaude O. J Cell Sci. 2010 Jun 15;123(Pt 12):2085-93.