TRIM25/EFP Rabbit Polyclonal Antibody, Unconjugated

Artikelnummer: BYT-ORB738470
Artikelname: TRIM25/EFP Rabbit Polyclonal Antibody, Unconjugated
Artikelnummer: BYT-ORB738470
Hersteller Artikelnummer: orb738470
Alternativnummer: BYT-ORB738470-100
Hersteller: Biorbyt
Wirt: Rabbit
Kategorie: Antikörper
Applikation: ELISA, FC, ICC, IF, IHC, WB
Spezies Reaktivität: Mouse, Rat
Immunogen: E.coli-derived mouse TRIM25/EFP recombinant protein (Position: M1-H590).
Konjugation: Unconjugated
Alternative Synonym: EFP, RING finger protein 147, RNF147, TRIM25, tripartite motif containing 25, Z147, Zinc finger protein 147, ZNF147
Anti-TRIM25/EFP Antibody. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Mouse, Rat.
Klonalität: Polyclonal
Konzentration: 500 µg/ml
Molekulargewicht: 71 kDa
UniProt: Q61510
Puffer: Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Formulierung: Lyophilized
Target-Kategorie: E3 ubiquitin/ISG15 ligase TRIM25
Application Verdünnung: Western blot, 0.25-0.5µg/ml, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5µg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5µg/ml, Mouse Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Mouse, Rat ELISA, 0.1-0.5µg/ml, -
Flow Cytometry analysis of NRK cells using anti-TRIM25/EFP antibody. Overlay histogram showing NRK cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRIM25/EFP Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Flow Cytometry analysis of RAW264.7 cells using anti-TRIM25/EFP antibody. Overlay histogram showing RAW264.7 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRIM25/EFP Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10 6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
IF analysis of TRIM25/EFP using anti-TRIM25/EFP antibody. TRIM25/EFP was detected in immunocytochemical section of MFC 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 5 µg/mL rabbit anti-TRIM25/EFP Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
IHC analysis of TRIM25/EFP using anti-TRIM25/EFP antibody. TRIM25/EFP 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 2 µg/ml rabbit anti-TRIM25/EFP Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
IHC analysis of TRIM25/EFP using anti-TRIM25/EFP antibody. TRIM25/EFP 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 2 µg/ml rabbit anti-TRIM25/EFP Antibody overnight at 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Western blot analysis of TRIM25/EFP using anti-TRIM25/EFP 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 30 ug of sample under reducing conditions. Lane 1: rat liver tissue lysates, Lane 2: rat RH35 whole cell lysates, Lane 3: mouse liver tissue lysates, Lane 4: mouse stomach tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA 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-TRIM25/EFP antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, the