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Product Info Summary
| SKU: | A00244-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-DDX58 Antibody Picoband®
SKU/Catalog Number
A00244-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-DDX58 Antibody Picoband® catalog # A00244-2. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
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.
Cite This Product
Anti-DDX58 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00244-2)
Host
Rabbit
Contents
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 method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human DDX58 recombinant protein (Position: H871-K925). Human DDX58 shares 83.3% amino acid (aa) sequence identity with mouse DDX58.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00244-2 is reactive to DDX58 in Human, Mouse, Rat
Observed Molecular Weight
107 kDa
Calculated molecular weight
106.6 kDa
Background of DDX58
RIG-I (retinoic acid-inducible gene I) is a RIG-I-like receptor dsRNA helicase enzyme that is encoded (in humans) by the DDX58 gene. RIG-I is part of the RIG-I-like receptor family, which also includes MDA5 and LGP2, and functions as a pattern recognition receptor that is a sensor for viruses such as influenza A, Sendai virus, and flavivirus. DEAD box proteins, characterized by the conserved motif Asp-Glu-Ala-Asp (DEAD), are putative RNA helicases which are implicated in a number of cellular processes involving RNA binding and alteration of RNA secondary structure. This gene encodes a protein containing RNA helicase-DEAD box protein motifs and a caspase recruitment domain (CARD). It is involved in viral double-stranded (ds) RNA recognition and the regulation of immune response.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00244-2 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human HepG2 whole cell, rat C6 whole cell, human Hela whole cell, mouse RAW2647 whole cell
ICC/IF: U20S cell
IF: human intestine cancer tissue
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of DDX58 using anti-DDX58 antibody (A00244-2).
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 (Catalog # A00244-2) 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 # EK1002) 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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IF analysis of DDX58 using anti-DDX58 antibody (A00244-2).
DDX58 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-DDX58 Antibody (A00244-2) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.
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IF analysis of DDX58 using anti-DDX58 antibody (A00244-2).
DDX58 was detected in a paraffin-embedded section of human intestine cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, 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 (A00244-2) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) was used as secondary antibody at 1:500 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.
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Flow Cytometry analysis of A431 cells using anti-DDX58 antibody (A00244-2).
Overlay histogram showing A431 cells stained with A00244-2 (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 (A00244-2,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-DDX58 Antibody Picoband® (A00244-2)
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1 Customer Q&As for Anti-DDX58 Antibody Picoband®
Question
We are currently using anti-DDX58 antibody A00244-2 for mouse tissue, and we are well pleased with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on pig tissues as well?
Verified Customer
Verified customer
Asked: 2017-10-19
Answer
The anti-DDX58 antibody (A00244-2) has not been tested for cross reactivity specifically with pig tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in pig you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-10-19
