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[KO Validated] IRF3 Rabbit mAb (A27272)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates using [KO Validated] IRF3 Rabbit mAb (A27272) at 1:11000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Western blot analysis of lysates from wild type (WT) and IRF3 knockout (KO) HeLa cells using [KO Validated] IRF3 Rabbit mAb (A27272) at 1:11000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

ABclonal:Immunohistochemistry - [KO Validated] IRF3 Rabbit mAb (A27272)

Immunohistochemistry analysis of paraffin-embedded Human breast cancer tissue using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human kidney tissue using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human thyroid cancer tissue using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse intestin tissue using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded HeLa and HeLa-IRF3-KO cells using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:600 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KO Validated] IRF3 Rabbit mAb (A27272)

Confocal imaging of NIH/3T3 cells using [KO Validated] IRF3 Rabbit mAb (A27272, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of HeLa-IRF3-KO cells and HeLa cells using [KO Validated] IRF3 Rabbit mAb (A27272, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - [KO Validated] IRF3 Rabbit mAb (A27272)

Immunoprecipitation of IRF3 from 300 µg extracts of HeLa cells was performed using 0.5 µg of [KO Validated] IRF3 Rabbit mAb (A27272). Rabbit Control IgG (AC005) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KO Validated] IRF3 Rabbit mAb (A27272) at a dilution of 1:10000.

Overview

Product name	[KO Validated] IRF3 Rabbit mAb
Catalog No.	A27272
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC73216

Background

This gene encodes a member of the interferon regulatory transcription factor (IRF) family. The encoded protein is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP. This complex translocates to the nucleus and activates the transcription of interferons alpha and beta, as well as other interferon-induced genes. The protein plays an important role in the innate immune response against DNA and RNA viruses. Mutations in this gene are associated with Encephalopathy, acute, infection-induced, herpes-specific, 7.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1-260 of human IRF3 (NP_001184051.1).
Sequence	MGTPKPRILPWLVSQLDLGQLEGVAWVNKSRTRFRIPWKHGLRQDAQQEDFGIFQAWAEATGAYVPGRDKPDLPTWKRNFRSALNRKEGLRLAEDRSKDPHDPHKIYEFVNSGVGDFSQPDTSPDTNGGGSTSDTQEDILDELLGNMVLAPLPDPGPPSLAVAPEPCPQPLRSPSLDNPTPFPNLGPSENPLKRLLVPGEEWEFEVTAFYRGRQVFQQTISCPEGLRLVGSEVGDRTLPGWPVTLPDPGMSLTDRGVMSY
Gene ID
Swiss Prot
Synonyms	IIAE7
Calculated MW	47kDa
Observed MW	45-55kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:11000 - 1:44000 IHC-P 1:600 - 1:3000 IF/ICC 1:100 - 1:400 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Immunoprecipitation
Positive samples	A-549, RAW 264.7, Rat spleen, HeLa
Cellular location	Cytoplasm, Nucleus.