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|
FCM (Flow Cytometry)
(Figure 10. Flow Cytometry analysis of A549 cells using anti- U2AF65/U2AF2 antibody (AAA19375).Overlay histogram showing A549 cells stained with AAA19375 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-U2AF65/U2AF2 Antibody (AAA19375, 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.)
IF (Immunofluorescence)
(Figure 9. IF analysis of U2AF65/U2AF2 using anti- U2AF65/U2AF2 antibody (AAA19375).U2AF65/U2AF2 was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti- U2AF65/U2AF2 Antibody (AAA19375) 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.)
IHC (Immunohistochemistry)
(Figure 8. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 7. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
IHC (Immunohistchemistry)
(Figure 6. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 5. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 4. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 3. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
IHC (Immunohistochemistry)
(Figure 2. IHC analysis of U2AF65/U2AF2 using anti U2AF65/U2AF2 antibody (AAA19375).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 (AAA19375) 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 # with DAB as the chromogen.)
WB (Western Blot)
(Figure 1. Western blot analysis of U2AF65/U2AF2 using anti-U2AF65/U2AF2 antibody (AAA19375).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 # AAA19375) 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 # 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.)
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.
