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Catalog Number | orb654378 |
---|---|
Category | Antibodies |
Description | U2AF65/U2AF2 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human U2AF65/U2AF2 recombinant protein (Position: M238-H470). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human, Mouse, Rat Direct ELISA, 0.1-0.5μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 65 kDa |
UniProt ID | P26368 |
Storage | Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles. |
Alternative names | Splicing factor U2AF 65 kDa subunit; U2 auxiliary Read more... |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of U2AF2 using anti-U2AF2 antibody (orb654378). 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 lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human U2OS whole cell lysates, Lane 4: rat brain tissue lysates, Lane 5: rat spleen tissue lysates, Lane 6: mouse brain tissue 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 rabbit anti-U2AF2 antigen affinity purified polyclonal antibody (Catalog # orb654378) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for U2AF2 at approximately 65KD. The expected band size for U2AF2 is at 65KD.
IHC analysis of U2AF2 using anti-U2AF2 antibody (orb654378). U2AF2 was detected in paraffin-embedded section of human mammary 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 1μg/ml rabbit anti-U2AF2 Antibody (orb654378) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IHC analysis of U2AF2 using anti-U2AF2 antibody (orb654378). U2AF2 was detected in paraffin-embedded section of mouse spleen 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 1μg/ml rabbit anti-U2AF2 Antibody (orb654378) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IHC analysis of U2AF2 using anti-U2AF2 antibody (orb654378). U2AF2 was detected in paraffin-embedded section of rat spleen 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 1μg/ml rabbit anti-U2AF2 Antibody (orb654378) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.
IF analysis of U2AF2 using anti-U2AF2 antibody (orb654378). U2AF2 was detected in immunocytochemical section of HeLa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-U2AF2 Antibody (orb654378) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of PC-3 cells using anti-U2AF2 antibody (orb654378). Overlay histogram showing PC-3 cells stained with orb654378 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-U2AF2 Antibody (orb654378, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Flow Cytometry analysis of ANA-1 cells using anti-U2AF2 antibody (orb654378). Overlay histogram showing ANA-1 cells stained with orb654378 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-U2AF2 Antibody (orb654378, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Flow Cytometry analysis of NRK cells using anti-U2AF2 antibody (orb654378). Overlay histogram showing NRK cells stained with orb654378 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-U2AF2 Antibody (orb654378, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
FC, ICC, IF, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
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