Western Blot (WB)
(Figure 1. Western blot analysis of U2AF65/U2AF2 using anti-U2AF65/U2AF2 antibody (MBS1754024).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HEK293 whole cell lysatesLane 2: human THP-1 whole cell lysatesLane 3: human U20S whole cell lysatesLane 4: human Jurkat whole cell lysatesLane 5: rat PC-12 whole cell lysatesLane 6: mouse brain tissue lysatesLane 7: rat brain tissue lysatesLane 8: mouse RAW264. 7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-U2AF65/U2AF2 antigen affinity purified monoclonal antibody (Catalog # MBS1754024) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for U2AF65/U2AF2 at approximately 65KD. The expected band size for U2AF65/U2AF2 is at 65KD. )
Mouse U2AF65/U2AF2 Monoclonal Antibody | anti-U2AF2 antibody
Anti-U2AF65/U2AF2 Antibody (monoclonal, 10F4)
IHC-P: 2-5ug/ml|Human, Mouse, Rat|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of U2AF65/U2AF2 using anti-U2AF65/U2AF2 antibody (MBS1754024).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HEK293 whole cell lysatesLane 2: human THP-1 whole cell lysatesLane 3: human U20S whole cell lysatesLane 4: human Jurkat whole cell lysatesLane 5: rat PC-12 whole cell lysatesLane 6: mouse brain tissue lysatesLane 7: rat brain tissue lysatesLane 8: mouse RAW264. 7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-U2AF65/U2AF2 antigen affinity purified monoclonal antibody (Catalog # MBS1754024) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for U2AF65/U2AF2 at approximately 65KD. The expected band size for U2AF65/U2AF2 is at 65KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of U2AF65/U2AF2 using anti-U2AF65/U2AF2 antibody (MBS1754024).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human HEK293 whole cell lysatesLane 2: human THP-1 whole cell lysatesLane 3: human U20S whole cell lysatesLane 4: human Jurkat whole cell lysatesLane 5: rat PC-12 whole cell lysatesLane 6: mouse brain tissue lysatesLane 7: rat brain tissue lysatesLane 8: mouse RAW264. 7 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-U2AF65/U2AF2 antigen affinity purified monoclonal antibody (Catalog # MBS1754024) at 0. 25 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for U2AF65/U2AF2 at approximately 65KD. The expected band size for U2AF65/U2AF2 is at 65KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human gallbladder adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human gallbladder adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human adnexal serous adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human adnexal serous adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 5. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human colonic adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 6. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 7. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 7. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 8. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 8. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunofluorescence (IF)
(Figure 9. IF analysis of U2AF65/U2AF2 using anti- U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 9. IF analysis of U2AF65/U2AF2 using anti- U2AF65/U2AF2 antibody (MBS1754024).U2AF65/U2AF2 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- U2AF65/U2AF2 Antibody (MBS1754024) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 10. Flow Cytometry analysis of A549 cells using anti- U2AF65/U2AF2 antibody (MBS1754024).Overlay histogram showing A549 cells stained with MBS1754024 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-U2AF65/U2AF2 Antibody (MBS1754024, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 10. Flow Cytometry analysis of A549 cells using anti- U2AF65/U2AF2 antibody (MBS1754024).Overlay histogram showing A549 cells stained with MBS1754024 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-U2AF65/U2AF2 Antibody (MBS1754024, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Mackereth, C. D., Madl, T., Bonnal, S., Simon, B., Zanier, K., Gasch, A., Rybin, V., Valcarcel, J., Sattler, M. Multi-domain conformational selection underlies pre-mRNA splicing regulation by U2AF. Nature 475: 408-411, 2011.
3. Sickmier, E. A., Frato, K. E., Shen, H., Paranawithana, S. R., Green, M. R., Kielkopf, C. L. Structural basis for polypyrimidine tract recognition by the essential pre-mRNA splicing factor U2AF65. Molec. Cell 23: 49-59, 2006.
NCBI and Uniprot Product Information
NCBI Description
U2 auxiliary factor (U2AF), comprised of a large and a small subunit, is a non-snRNP protein required for the binding of U2 snRNP to the pre-mRNA branch site. This gene encodes the U2AF large subunit which contains a sequence-specific RNA-binding region with 3 RNA recognition motifs and an Arg/Ser-rich domain necessary for splicing. The large subunit binds to the polypyrimidine tract of introns early during spliceosome assembly. Multiple transcript variants have been detected for this gene, but the full-length natures of only two have been determined to date. [provided by RefSeq, Jul 2008]
Uniprot Description
U2AF2: Necessary for the splicing of pre-mRNA. 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. Belongs to the splicing factor SR family. 2 isoforms of the human protein are produced by alternative splicing.
Protein type: RNA-binding; Spliceosome; RNA splicing
Chromosomal Location of Human Ortholog: 19q13.42
Cellular Component: nucleoplasm; spliceosome; nuclear speck
Molecular Function: protein binding; enzyme binding; nucleotide binding
Biological Process: transcription from RNA polymerase II promoter; nuclear mRNA splicing, via spliceosome; mRNA export from nucleus; negative regulation of nuclear mRNA splicing, via spliceosome; RNA splicing; gene expression; mRNA 3'-end processing; mRNA processing; termination of RNA polymerase II transcription; positive regulation of RNA splicing
Research Articles on U2AF2
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Product Notes
The U2AF2 u2af2 (Catalog #AAA1754024) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-U2AF65/U2AF2 Antibody (monoclonal, 10F4) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's U2AF65/U2AF2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM). WB: 0.1-0.25ug/ml|Human, Mouse, Rat| IHC-P: 2-5ug/ml|Human, Mouse, Rat| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the U2AF2 u2af2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "U2AF65/U2AF2, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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