Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3. *This antibody is supplied in a stabilized formulation. Compatibility with conjugation reactions depends on the chemistry of the conjugation
Form:
Lyophilized
Target:
Antiviral innate immune response receptor RIG-I
Application Dilute:
Western blot, 0.1-0.5µg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2µg/ml, Human Immunofluorescence, 5µg/ml, Human Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Human
Flow Cytometry analysis of A431 cells using anti-DDX58 antibody. Overlay histogram showing A431 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-DDX58 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 A431 cells using anti-DDX58 antibody (Blu
IF analysis of DDX58 using anti-DDX58 antibody. DDX58 was detected in a paraffin-embedded section of human intestine 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 5 µg/mL rabbit anti-DDX58 Antibody overnight at 4C. DyLight550 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 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.
IF analysis of DDX58 using anti-DDX58 antibody. DDX58 was detected in immunocytochemical section of U20S 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 2 µg/mL rabbit anti-DDX58 Antibody overnight at 4C. DyLight594 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.
Western blot analysis of DDX58 using anti-DDX58 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: human HepG2 whole cell lysates, Lane 2: rat C6 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: mouse RAW264.7 whole cell 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-DDX58 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 with Tanon 5200 system. A specific band was detected for DDX58 at approximately 107 kDa. The expected band size for DDX58 is at 107 kDa.
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