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Phospho-IRF3 (S386) Recombinant Rabbit Monoclonal Antibody [SU03-28]

Usd: 385

Specification

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Catalog# ET1608-22

Phospho-IRF3 (S386) Recombinant Rabbit Monoclonal Antibody [SU03-28]

Application

  • WB

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-IRF3 (S386) Recombinant Rabbit Monoclonal Antibody [SU03-28]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser386 of human IRF3.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 47 kDa

Positive Control

MCF7 treated with 100nM Calyculin A for 30 minutes whole cell lysate, MCF-7.

Conjugation

unconjugated

Clone Number

SU03-28

RRID

AB_3069793

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

Target

Function

Interferon regulatory factor 3, also known as IRF3, is an interferon regulatory factor. IRF3 is a member of the interferon regulatory transcription factor (IRF) family. IRF3 was originally discovered as a homolog of IRF1 and IRF2. IRF3 has been further characterized and shown to contain several functional domains including a nuclear export signal, a DNA-binding domain, a C-terminal IRF association domain and several regulatory phosphorylation sites. IRF3 is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP. The complex translocates into the nucleus for the transcriptional activation of interferons alpha and beta, and further interferon-induced genes.

Background References

1. Wheeler LA et al. TREX1 Knockdown Induces an Interferon Response to HIV that Delays Viral Infection in Humanized Mice. Cell Rep 15:1715-27 (2016).

2. Meng X et al. C7L family of poxvirus host range genes inhibits antiviral activities induced by type I interferons and interferon regulatory factor 1. J Virol 86:4538-47 (2012).

Sequence Similarity

Belongs to the IRF family.

Tissue Specificity

Expressed constitutively in a variety of tissues.

Post-translational Modification

Constitutively phosphorylated on many Ser/Thr residues. Activated following phosphorylation by TBK1 and IKBKE. Innate adapter protein MAVS, STING1 or TICAM1 are first activated by viral RNA, cytosolic DNA, and bacterial lipopolysaccharide (LPS), respectively, leading to activation of the kinases TBK1 and IKBKE. These kinases then phosphorylate the adapter proteins on the pLxIS motif, leading to recruitment of IRF3, thereby licensing IRF3 for phosphorylation by TBK1. Phosphorylated IRF3 dissociates from the adapter proteins, dimerizes, and then enters the nucleus to induce IFNs.; (Microbial infection) Phosphorylation and subsequent activation of IRF3 is inhibited by vaccinia virus protein E3.; Ubiquitinated; ubiquitination involves RBCK1 leading to proteasomal degradation. Polyubiquitinated; ubiquitination involves TRIM21 leading to proteasomal degradation.; ISGylated by HERC5 resulting in sustained IRF3 activation and in the inhibition of IRF3 ubiquitination by disrupting PIN1 binding. The phosphorylation state of IRF3 does not alter ISGylation.

Subcellular Location

Cytoplasm, Nucleus, Mitochondrion.

UNIPROT

SwissProt: Q14653 Human

Synonyms

IIAE7 antibody

Interferon regulatory factor 3 antibody

IRF 3 antibody

IRF-3 antibody

IRF3 antibody

IRF3_HUMAN antibody

MGC94729 antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-IRF3 (S386) on different lysates with Rabbit anti-Phospho-IRF3 (S386) antibody (<a href="/products/ET1608-22" style="font-weight: bold;text-decoration: underline;">ET1608-22</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7 whole cell lysate<br />Lane 2: MCF7 treated with 100nM Calyculin A for 30 minutes whole cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 47 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 5 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-22" style="font-weight: bold;text-decoration: underline;">ET1608-22</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-IRF3 (S386) on different lysates with Rabbit anti-Phospho-IRF3 (S386) antibody (ET1608-22) at 1/1,000 dilution.

    Lane 1: MCF7 whole cell lysate
    Lane 2: MCF7 treated with 100nM Calyculin A for 30 minutes whole cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 47 kDa
    Observed band size: 55 kDa

    Exposure time: 5 minutes;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-22) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • ICC staining of Phospho-IRF3 (S386) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1608-22" style="font-weight: bold;text-decoration: underline;">ET1608-22</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Phospho-IRF3 (S386) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1608-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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