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FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of K562 cells using anti-IRF3 antibody (AAA19216).Overlay histogram showing K562 cells stained with AAA19216 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IRF3 Antibody (AAA19216, 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.)

Rabbit anti-Human IRF3 Polyclonal Antibody | anti-IRF3 antibody

Anti-IRF3 Antibody

Reactivity
Human
Applications
Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
IRF3; Polyclonal Antibody; Anti-IRF3 Antibody; Interferon regulatory factor 3; IRF-3; interferon regulatory factor 3; anti-IRF3 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for IRF3 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Applicable Applications for anti-IRF3 antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml|Human|
IHC-P: 2-5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Immunogen
E Coli-derived human IRF3 recombinant protein (Position: K5-Q422).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 6. Flow Cytometry analysis of K562 cells using anti-IRF3 antibody (AAA19216).Overlay histogram showing K562 cells stained with AAA19216 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IRF3 Antibody (AAA19216, 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 6. Flow Cytometry analysis of K562 cells using anti-IRF3 antibody (AAA19216).Overlay histogram showing K562 cells stained with AAA19216 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IRF3 Antibody (AAA19216, 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 CACO-2 cells using anti-IRF3 antibody (AAA19216).Overlay histogram showing CACO-2 cells stained with AAA19216 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IRF3 Antibody (AAA19216, 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 CACO-2 cells using anti-IRF3 antibody (AAA19216).Overlay histogram showing CACO-2 cells stained with AAA19216 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IRF3 Antibody (AAA19216, 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.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of IRF3 using anti-IRF3 antibody (AAA19216).IRF3 was detected in paraffin-embedded section of human rectal 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 2μg/ml rabbit anti-IRF3 Antibody (AAA19216) 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 4. IHC analysis of IRF3 using anti-IRF3 antibody (AAA19216).IRF3 was detected in paraffin-embedded section of human rectal 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 2μg/ml rabbit anti-IRF3 Antibody (AAA19216) 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 3. IHC analysis of IRF3 using anti-IRF3 antibody (AAA19216).IRF3 was detected in paraffin-embedded section of human mammary 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 2μg/ml rabbit anti-IRF3 Antibody (AAA19216) 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 3. IHC analysis of IRF3 using anti-IRF3 antibody (AAA19216).IRF3 was detected in paraffin-embedded section of human mammary 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 2μg/ml rabbit anti-IRF3 Antibody (AAA19216) 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 IRF3 using anti-IRF3 antibody (AAA19216).IRF3 was detected in paraffin-embedded section of human lung 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 2μg/ml rabbit anti-IRF3 Antibody (AAA19216) 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 IRF3 using anti-IRF3 antibody (AAA19216).IRF3 was detected in paraffin-embedded section of human lung 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 2μg/ml rabbit anti-IRF3 Antibody (AAA19216) 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 IRF3 using anti-IRF3 antibody (AAA19216).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 50ug of sample under reducing conditions.Lane 1: human PC-3 whole cell 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 # AAA19216) 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 # Tanon 5200 system. A specific band was detected for IRF3 at approximately 55KD. The expected band size for IRF3 is at 55KD.)

WB (Western Blot) (Figure 1. Western blot analysis of IRF3 using anti-IRF3 antibody (AAA19216).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 50ug of sample under reducing conditions.Lane 1: human PC-3 whole cell 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 # AAA19216) 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 # Tanon 5200 system. A specific band was detected for IRF3 at approximately 55KD. The expected band size for IRF3 is at 55KD.)
Related Product Information for anti-IRF3 antibody
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 with CREBBP. IRF3 plays an important role in the innate immune system's response to viral infection. Aggregated MAVS have been found to activate IRF3 dimerization. Although IRF3 increased transcriptional activity from an ISRE-containing promoter, expression of IRF3 as 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 with IRF3 only subsequent to viral infection, and the authors stated that these are also subunits of DRAF1.
References
1. Bellingham, J., Gregory-Evans, K., Gregory-Evans, C. Y. Mapping of human interferon regulatory factor 3 (IRF3) to chromosome 19q13. 3-13. 4 by an intragenic polymorphic marker. Ann. Hum. Genet. 62: 231-234, 1998.
2. Doyle, S. E., Vaidya, S. A., O'Connell, R., Dadgostar, H., Dempsey, P. W., Wu, T. -T., Rao, G., Sun, R., Haberland, M. E., Modlin, R. L., Cheng, G. IRF3 mediates a TLR3/TLR4-specific antiviral gene program. Immunity 17: 251-263, 2002.
3. Stetson, D. B., Medzhitov, R. Recognition of cytosolic DNA activates an IRF3-dependent innate immune response. Immunity 24: 93-103, 2006.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
47,219 Da
NCBI Official Full Name
interferon regulatory factor 3 isoform 4
NCBI Official Synonym Full Names
interferon regulatory factor 3
NCBI Official Symbol
IRF3
NCBI Protein Information
interferon regulatory factor 3
UniProt Protein Name
Interferon regulatory factor 3
UniProt Gene Name
IRF3
UniProt Synonym Gene Names
IRF-3
UniProt Entry Name
IRF3_HUMAN

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Product Notes

The IRF3 irf3 (Catalog #AAA19216) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-IRF3 Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's IRF3 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human| IHC-P: 2-5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the IRF3 irf3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "IRF3, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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