Product Info Summary
SKU: | A00244-3 |
---|---|
Size: | 100 µg/vial |
Reactive Species: | Human |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-DDX58/RIG-1 Antibody Picoband™
SKU/Catalog Number
A00244-3
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-DDX58/RIG-1 Antibody Picoband™ catalog # A01000-5. Tested in WB, IHC, FCM, ELISA applications. This antibody reacts with Human.
Storage & Handling
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 freezing and thawing.
Cite This Product
Anti-DDX58/RIG-1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00244-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Clone Number
1B9
Isotype
IgG
Immunogen
E.coli-derived human DDX58/RIG-1 recombinant protein (Position: K203-K925). Human RIGI shares 78.2% amino acid (aa) sequence identity with mouse RIGI.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
A00244-3 is reactive to DDX58 in Human
Applications
A00244-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee
Observed Molecular Weight
101 kDa
Calculated molecular weight
106.6kDa
Background of RIG-I
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 the antiviral innate immune response. Mutations in this gene are associated with Singleton-Merten syndrome 2.
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.
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Assay dilution & Images
Reconsitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
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, 0.25-0.5 µg/ml, Human
Immunohistochemistry, 2-5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x106 cells, Human
ELISA, 0.1-0.5 µg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
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 A431 whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human Hela 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/RIG-1 antigen affinity purified polyclonal antibody (Catalog # A00244-3) 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/RIG-1 at approximately 101 kDa. The expected band size for DDX58/RIG-1 is at 107 kDa.
Click image to see more details
Figure 2. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human appendicitis 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human appendicitis 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human cervix squamous cell carcinoma 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human cervix squamous cell carcinoma 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human rectum adenocarcinoma 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 7. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human rectum adenocarcinoma 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 8. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human spleen 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 9. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human spleen 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 10. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human testicular seminoma 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 11. IHC analysis of DDX58/RIG-1 using anti-DDX58/RIG-1 antibody (A00244-3).
DDX58/RIG-1 was detected in a paraffin-embedded section of human testicular seminoma 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 2 μg/ml rabbit anti-DDX58/RIG-1 Antibody (A00244-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 12. Flow Cytometry analysis of MCF-7 cells using anti-DDX58/RIG-1 antibody (A00244-3).
Overlay histogram showing MCF-7 cells stained with A00244-3 (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/RIG-1 Antibody (A00244-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For DDX58 (Source: Uniprot.org, NCBI)
Gene Name
DDX58
Full Name
Antiviral innate immune response receptor RIG-I
Weight
106.6kDa
Superfamily
helicase family
Alternative Names
DDX58; DEAD (Asp-Glu-Ala-Asp) box polypeptide 58; DEAD box protein 58; DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide; DKFZp686N19181; EC 3.6.4.13; FLJ13599; probable ATP-dependent RNA helicase DDX58; retinoic acid inducible gene I; Retinoic acid-inducible gene 1 protein; Retinoic acid-inducible gene I protein; RIG-1; RIGI; RIG-I; RIG-IDKFZp434J1111; RNA helicase RIG-I DDX58 RIG-I, RIG1, RIGI, RLR-1, SGMRT2 DExD/H-box helicase 58 antiviral innate immune response receptor RIG-I|DEAD (Asp-Glu-Ala-Asp) box polypeptide 58|DEAD box protein 58|DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide|RNA helicase RIG-I|probable ATP-dependent RNA helicase DDX58|retinoic acid-inducible gene 1 protein|retinoic acid-inducible gene I protein
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on DDX58, check out the DDX58 Infographic
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In this infographic, you will see the following information for DDX58: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-DDX58/RIG-1 Antibody Picoband™ (A00244-3)
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