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IRF9 Recombinant Rabbit Monoclonal Antibody [PSH08-20]

Usd: 385

Specification

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Catalog# HA722963

IRF9 Recombinant Rabbit Monoclonal Antibody [PSH08-20]

Application

  • WB

  • IF-Cell

  • IP

  • ChIP

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

IRF9 Recombinant Rabbit Monoclonal Antibody [PSH08-20]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human IRF9 aa 1-393.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IP, ChIP

Molecular Weight

Predicted band size: 44 kDa

Positive Control

HeLa cell lysate, HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate, Daudi cell lysate, Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate, Jurkat cell lysate, Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate, THP-1 cell lysate, HeLa cells treated with 10ng/mL IFN-α1 for 16 hours.

Conjugation

unconjugated

Clone Number

PSH08-20

Reactivity Data

WB IF-Cell IP ChIP
Human
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Mouse
Not Recommended
Rat
Not Recommended

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IP

  • 1-2μg/sample

  • ChIP

  • Use 2~5 μg for 25 μg of chromatin.

Target

Function

Interferon regulatory factor 9 is a protein that in humans is encoded by the IRF9 gene, previously known as ISGF3G. Transcription factor that plays an essential role in anti-viral immunity. It mediates signaling by type I IFNs (IFN-alpha and IFN-beta). Following type I IFN binding to cell surface receptors, Jak kinases (TYK2 and JAK1) are activated, leading to tyrosine phosphorylation of STAT1 and STAT2. IRF9/ISGF3G associates with the phosphorylated STAT1:STAT2 dimer to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of interferon stimulated genes, which drive the cell in an antiviral state.

Background References

1. Lan C et al. Suppression of IRF9 Promotes Osteoclast Differentiation by Decreased Ferroptosis via STAT3 Activation. Inflammation. 2024 Feb

2. Zhao Q et al. An MRTF-A-ZEB1-IRF9 axis contributes to fibroblast-myofibroblast transition and renal fibrosis. Exp Mol Med. 2023 May

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q00978 Human

Synonyms

IFN alpha responsive transcription factor subunit antibody

IFN-alpha-responsive transcription factor subunit antibody

Interferon regulatory factor 9 antibody

interferon stimulated transcription factor 3 antibody

Interferon-stimulated gene factor 3 gamma antibody

interferon-stimulated transcription factor 3, gamma 48kDa antibody

IRF 9 antibody

IRF-9 antibody

Irf9 antibody

IRF9_HUMAN antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody (<a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate<br />Lane 3: Daudi cell lysate<br />Lane 4: Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate<br />Lane 5: Jurkat cell lysate<br />Lane 6: Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate<br />Lane 7: THP-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 44 kDa<br />Observed band size: 48 kDa<br /><br />Exposure time: 46 seconds; ECL: K1801;<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/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody (HA722963) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 10ng/mL IFN-α1 for 16 hours cell lysate
    Lane 3: Daudi cell lysate
    Lane 4: Daudi treated with 10ng/mL IFN-α1 for 16 hours cell lysate
    Lane 5: Jurkat cell lysate
    Lane 6: Jurkat treated with 100nM IFN-α1 for 24 hours cell lysate
    Lane 7: THP-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 44 kDa
    Observed band size: 48 kDa

    Exposure time: 46 seconds; ECL: K1801;

    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 (HA722963) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody (<a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a>) at 1/1,000 dilution.<br /><br />Lane 1: Human IRF9 recombinant protein, 30ng/Lane<br />Lane 2: Human IRF8 recombinant protein, 30ng/Lane<br /><br />Exposure time: 5 seconds; ECL: K1801;<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/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of IRF9 on different lysates with Rabbit anti-IRF9 antibody (HA722963) at 1/1,000 dilution.

    Lane 1: Human IRF9 recombinant protein, 30ng/Lane
    Lane 2: Human IRF8 recombinant protein, 30ng/Lane

    Exposure time: 5 seconds; ECL: K1801;

    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 (HA722963) 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 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9 with Rabbit anti-IRF9 antibody (<a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IRF9 antibody (<a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with 10ng/mL IFN-α1 for 16 hours labeling IRF9 with Rabbit anti-IRF9 antibody (HA722963) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IRF9 antibody (HA722963) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • IRF9 was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa cell lysate (input)<br />Lane 2: <a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    IRF9 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722963 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722963 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA722963 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA722963 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute; ECL: K1802

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 10ng/mL IFN-α1 for 16 hours with IRF9 (<a href="/products/HA722963" style="font-weight: bold;text-decoration: underline;">HA722963</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    ☑ Cell treatment (CT)

    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 10ng/mL IFN-α1 for 16 hours with IRF9 (HA722963) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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