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Product Info Summary
| SKU: | A00165-6 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-IRF3 Antibody Picoband®
SKU/Catalog Number
A00165-6
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-IRF3 Antibody Picoband® catalog # A00165-6. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse. 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-IRF3 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00165-6)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human IRF3 recombinant protein (Position: M1-S427).
*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
A00165-6 is reactive to IRF3 in Human, Mouse
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
55 kDa
Calculated molecular weight
47.219kDa
Background of IRF3
IRF3 (interferon regulatory factor 3)is a member of the interferon regulatory transcription factor (IRF) family. The IRF3 gene is mapped on 19q13.33. IRF3 is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex withCREBBP. IRF3 plays an important role in theinnate immune system's response toviral infection. AggregatedMAVShave been found to activate IRF3 dimerization. AlthoughIRF3increased transcriptional activity from an ISRE-containing promoter, expression ofIRF3as a Gal4 fusion protein did not activate expression of a chloramphenicol acetyltransferase (CAT) reporter gene containing repeats of the Gal4-binding sites.Translocation of IRF3 was accompanied by an increase in serine and threonine phosphorylation.The transcriptional activators CREBBP and EP300 coimmunoprecipitated withIRF3only subsequent to viral infection, and the authors stated that these are also subunits of DRAF1.
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
A00165-6 is guaranteed for ELISA, Flow Cytometry, IF, IHC, 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, 0.25-0.5μg/ml, Human, Mouse
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell, human Raji whole cell, human HepG2 whole cell, mouse lung tissue
IHC: human rectal cancer tissue
ICC/IF: A431 cell
FCM: K562 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of IRF3 using anti-IRF3 antibody (A00165-6).
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: human Hela whole cell lysates,
Lane 2: human Raji whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: mouse lung 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-IRF3 antigen affinity purified polyclonal antibody (Catalog # A00165-6) 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 IRF3 at approximately 55KD. The expected band size for IRF3 is at 55KD.
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Figure 2. IF analysis of IRF3 using anti-IRF3 antibody (A00165-6).
IRF3 was detected in immunocytochemical section of A431 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 5μg/mL rabbit anti-IRF3 Antibody (A00165-6) 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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Figure 3. Flow Cytometry analysis of K562 cells using anti-IRF3 antibody (A00165-6).
Overlay histogram showing K562 cells stained with A00165-6 (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-IRF3 Antibody (A00165-6, 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.
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Figure 4. IHC analysis of IRF3 using anti-IRF3 antibody (A00165-6).
IRF3 was detected in a paraffin-embedded section of human rectal 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 2 μg/ml rabbit anti-IRF3 Antibody (A00165-6) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For IRF3 (Source: Uniprot.org, NCBI)
Gene Name
IRF3
Full Name
Interferon regulatory factor 3
Weight
47.219kDa
Superfamily
IRF family
Alternative Names
Interleukin-17C; Il-17c; Cytokine CX2; Il17c IRF3 IIAE7 interferon regulatory factor 3 interferon regulatory factor 3
*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 IRF3, check out the IRF3 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for IRF3: 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-IRF3 Antibody Picoband® (A00165-6)
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