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AB258472

Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate

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IRF9 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and 2 bp deletion in exon2.
5 Images
Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)
  • WB

Lab

Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)

Lane 1 : Wild-type HeLa treated hIFN-a1 (10 ng/ml, 16 h) cell lysate 20 μg
Lane 2 : IRF9 knockout HeLa treated hIFN-a1 (10 ng/ml, 16 h) cell lysate 20 μg
Lane 3 : Wild-type HeLa vehicle control hIFN-a1 (0 ng/ml, 16 h) cell lysate 20 μg
Lane 4 : IRF9 knockout HeLa vehicle control hIFN-a1 (0 ng/ml, 16 h) cell lysate 20 μg
False colour image of Western blot : Anti-Interferon regulatory factor 9/IRF-9 antibody [14HCLC] staining at 2 μg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab277803 was shown to bind specifically to Interferon regulatory factor 9/IRF-9. A band was observed at 48 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in IRF9 knockout cell line ab266051 (knockout cell lysate ab258472). The band observed in the knockout lysate lane below 48 kDa is likely to represent a truncated form of Interferon regulatory factor 9/IRF-9. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and IRF9 knockout HeLa cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^®^ 20 (TBS-T)
before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^®^ 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®^ 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [14HCLC] (<a href='/en-us/products/primary-antibodies/interferon-regulatory-factor-9-irf-9-antibody-14hclc-ab277803'>ab277803</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa treated hIFN-a1 (10 ng/ml, 16 h) cell lysate at 20 µg

Lane 2:

IRF9 knockout HeLa treated hIFN-a1 (10 ng/ml, 16 h) cell lysate at 20 µg

Lane 3:

Wild-type HeLa vehicle control hIFN-a1 (0 ng/ml, 16 h) cell lysate at 20 µg

Lane 4:

IRF9 knockout HeLa vehicle control hIFN-a1 (0 ng/ml, 16 h) cell lysate at 20 µg

Predicted band size: 44 kDa

Observed band size: 48 kDa

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Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)
  • WB

Lab

Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)

Lane 1 : wild-type HeLa Treated Interferon-alpha1 (hIFN-alpha1) (10 ng/ml, 16 h) cell lysate 20 ug
Lane 2 : IRF9 knockout HeLa treated : hIFN-alpha1 (10 ng/ml, 16 h) cell lysate 20 ug
Lane 3 : wild-type HeLa Control Interferon-alpha1 (hIFN-alpha1) (0 ng/ml, 16 h) cell lysate 20 ug
Lane 4 : IRF9 knockout HeLa vehicle control : hIFN-alpha1 (0 ng/ml, 16 h) cell lysate 20 ug
Lane 5 : THP-1 cell lysate 20 ug
Lane 6 : ACHN cell lysate 20 ug
False colour image of Western blot : Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab271043 was shown to bind specifically to Interferon regulatory factor 9/IRF-9. A band was observed at 48 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in IRF9 knockout cell line ab266051 (knockout cell lysate ab258472). The band observed in the knockout lysate lane below 48 kDa is likely to represent a truncated form of Interferon regulatory factor 9/IRF-9. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and IRF9 knockout HeLa cell lysates were analysed.First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (<a href='/en-us/products/primary-antibodies/interferon-regulatory-factor-9-irf-9-antibody-epr24260-55-ab271043'>ab271043</a>) at 1/1000 dilution

Lane 1:

wild-type HeLa Treated Interferon-alpha1 (hIFN-a1) (10 ng/ml, 16 h) cell lysate at 20 µg

Lane 2:

IRF9 knockout HeLa treated: hIFN-a1 (10 ng/ml, 16 h) cell lysate at 20 µg

Lane 3:

wild-type HeLa Control Interferon-alpha1 (hIFN-a1) (0 ng/ml, 16 h) cell lysate at 20 µg

Lane 4:

IRF9 knockout HeLa vehicle control: hIFN-a1 (0 ng/ml, 16 h) cell lysate at 20 µg

Lane 5:

THP-1 cell lysate at 20 µg

Lane 6:

ACHN cell lysate at 20 µg

Predicted band size: 44 kDa

Observed band size: 48 kDa

false

Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)
  • WB

Lab

Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : IRF9 knockout HeLa cell lysate 20 μg
Lane 3 : THP-1 cell lysate 20 μg
Lane 4 : ACHN cell lysate 20 μg

Lanes 1 - 4 : Merged signal (red and green). Green - Anti-IRF9 antibody observed at 48 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa. Anti-IRF9 antibody was shown to react with IRF-9 in wild-type HeLa cells in western blot. The bands observed in IRF9 knockout cell line ab266051 (IRF9 knockout cell lysate ab258472) below 48 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and IRF9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with anti-IRF9 antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Anti-IRF9 antibody at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

IRF9 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-irf9-interferon-regulatory-factor-9-knockout-hela-cell-line-ab266051'>ab266051</a>)

Lane 3:

THP-1 cell lysate at 20 µg

Lane 4:

ACHN cell lysate at 20 µg

false

Sanger Sequencing - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)
  • Sanger seq

Unknown

Sanger Sequencing - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)

Allele-1 : 1 bp insertion in exon2

Sanger Sequencing - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)
  • Sanger seq

Unknown

Sanger Sequencing - Human IRF9 (Interferon regulatory factor 9) knockout HeLa cell lysate (AB258472)

Allele-2 : 2 bp deletion in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and 2 bp deletion in exon2.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB271043

Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Interferon regulatory factor 9/IRF-9 antibody [EPR24260-55] (AB271043)

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Cell Lines & Lysates

AB274943

Human IRF9 (Interferon regulatory factor 9) knockout MCF7 cell line

Next Generation Sequencing - Human IRF9 (Interferon regulatory factor 9) knockout MCF7 cell line (AB274943)

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  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
IRF9
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Interferon regulatory factor 9 (IRF-9) also known as ISGF3-γ is a critical component in the transcriptional regulation of interferon-stimulated genes. This protein with a mass of approximately 48 kDa serves as an important mediator in the interferon signaling pathway. IRF-9 is widely expressed in various tissues where it functions as part of the immune response to pathogens. Its expression can be induced by interferons signaling molecules involved in antiviral defense.
Biological function summary

IRF-9 participates in the assembly of the transcription factor complex ISGF3 alongside STAT1 and STAT2. This complex translocates into the nucleus to initiate the transcription of interferon-stimulated genes that promote antiviral states. IRF-9 through this functional role impacts a range of cellular responses including proliferation apoptosis and immune regulation. It affects the temporal and spatial aspects of gene expression in response to interferons.

Pathways

IRF-9 plays a significant role in both the Jak-STAT and interferon signaling pathways. These pathways are essential for the activation of genes involved in immune defense. Through its involvement in these pathways IRF-9 interacts with proteins such as STAT1 and STAT2 creating a bridge between extracellular signals and gene activation. This involvement ensures that cells can respond effectively to viral infections enhancing innate and adaptive immune responses.

IRF-9 has been implicated in autoimmune diseases and certain cancers. Dysfunction in the IRF-9 associated pathways can lead to conditions such as systemic lupus erythematosus where inefficient regulation of immune responses occurs. In cancer altered IRF-9 expression can contribute to uncontrolled cell proliferation. These connections highlight the importance of IRF-9 in maintaining cellular homeostasis and immune integrity. IRF-9's interaction with STAT proteins and interferon pathways highlights its central role in these disease processes.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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