Rabbit anti-Mouse, Rat TRIM25/EFP Polyclonal Antibody | anti-Trim25 antibody
Anti-TRIM25/EFP Antibody
IHC-P: 2-5ug/ml|Mouse, Rat|
ICC/IF: 5ug/ml|Mouse|
FC/FACS/FCM: 1-3ug/1x106 cells|Mouse, Rat|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
FCM (Flow Cytometry)
(Figure 6. Flow Cytometry analysis of RAW264. 7 cells using anti-TRIM25/EFP antibody (AAA19272).Overlay histogram showing RAW264. 7 cells stained with AAA19272 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRIM25/EFP Antibody (AAA19272, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
FCM (Flow Cytometry)
(Figure 5. Flow Cytometry analysis of NRK cells using anti-TRIM25/EFP antibody (AAA19272).Overlay histogram showing NRK cells stained with AAA19272 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRIM25/EFP Antibody (AAA19272, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)
IF (Immunofluorescence)
(Figure 4. IF analysis of TRIM25/EFP using anti- TRIM25/EFP antibody (AAA19272).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 5μg/mL rabbit anti- TRIM25/EFP Antibody (AAA19272) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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 3. IHC analysis of TRIM25/EFP using anti-TRIM25/EFP antibody (AAA19272).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 2μg/ml rabbit anti-TRIM25/EFP Antibody (AAA19272) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 TRIM25/EFP using anti-TRIM25/EFP antibody (AAA19272).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 2μg/ml rabbit anti-TRIM25/EFP Antibody (AAA19272) overnight at 4 degree C. Biotinylated goat anti-rabbit 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 TRIM25/EFP using anti-TRIM25/EFP antibody (AAA19272).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 30ug of sample under reducing conditions.Lane 1: rat liver tissue lysatesLane 2: rat RH35 whole cell lysatesLane 3: mouse liver tissue lysatesLane 4: mouse stomach 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-TRIM25/EFP antigen affinity purified polyclonal antibody (Catalog # AAA19272) at 0. 5 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-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 # with Tanon 5200 system. A specific band was detected for TRIM25/EFP at approximately 71KD. The expected band size for TRIM25/EFP is at 71KD.)
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