EWSR1
| Cat Number: | AB-84177 |
|---|---|
| Size: | 100 ug |
| Clone: | POLY |
| Concentration: | 1mg/ml |
| Host: | Rb |
| Isotype: | IgG |
| Immunogen: | A synthetic peptide corresponding to a sequence in the middle region of human EWSR1. (369-399aa NDSVTLDDLADFFKQCGVVKMNKRTGQPMIH), different from the related mouse sequence by one amino acid. |
| Reactivity: | Hu, Ms, Rt |
| Applications: | Western blot: 1:2000-1:10.000 |
| Purification: | Aff. Pur. |
| Background: | This gene encodes a multifunctional protein that is involved in various cellular processes, including gene expression, cell signaling, and RNA processing and transport. The protein includes an N-terminal transcriptional activation domain and a C-terminal RNA-binding domain. Chromosomal translocations between this gene and various genes encoding transcription factors result in the production of chimeric proteins that are involved in tumorigenesis. These chimeric proteins usually consist of the N-terminal transcriptional activation domain of this protein fused to the C-terminal DNA-binding domain of the transcription factor protein. Mutations in this gene, specifically a t(11;22)(q24;q12) translocation, are known to cause Ewing sarcoma as well as neuroectodermal and various other tumors. Alternative splicing of this gene results in multiple transcript variants. Related pseudogenes have been identified on chromosomes 1 and 14. |
| Form: | Liquid |
| Buffer: | Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3. |
| Storage: | At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing. |
Western blot analysis of EWSR1 using anti-EWSR1 antibody 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: Rat skeletal muscle Tissue Lysate,
Lane 2: Mouse skeletal muscle Tissue Lysate,
Lane 3: Mouse HEPA1-6 Whole Cell Lysate.
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-EWSR1 antigen affinity purified polyclonal antibody 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with
Tanon 5200 system. A specific band was detected for EWSR1 at approximately 90KD. The expected band size for EWSR1 is at
68KD.
IHC analysis of EWSR1 using anti-EWSR1 antibodyEWSR1 was detected in paraffin-embedded section of Mouse Testis Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins.
The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EWSR1 Antibody 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) with DAB as the chromogen.
IF analysis of EWSR1 using anti- EWSR1 antibody EWSR1 was detected in immunocytochemical section of U20S cell. 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 2μg/mL
rabbit anti- EWSR1 Antibody 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. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Flow Cytometry analysis of U20S cells using anti- EWSR1 antibody.
Overlay histogram showing U20S cells stained with Blue line).The cells were blocked with 10% normal goat serum.
And then incubated with rabbit anti-EWSR1 Antibody (1μg/1×106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1×106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1×106) used under the same conditions. Unlabelled sample (Red line) was
also used as a control.
Western blot analysis of EWSR1 using anti-EWSR1
antibody
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 Hela Whole Cell Lysate,
Lane 2: Human K562 Whole Cell Lysate,
Lane 3: Human Jurkat Whole Cell Lysate,
Lane 4: Human HL-60 Whole Cell Lysate,
Lane 5: human HEK293 Whole Cell Lysate,
Lane 6: Human Caco-2 Whole Cell Lysate.
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-EWSR1 antigen affinity purified polyclonal antibody 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced
Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for EWSR1 at approximately 90KD. The expected band size for EWSR1 is at 68KD.