AB263784

Human IRF3 knockout HeLa cell lysate

Name: Human IRF3 knockout HeLa cell lysate (ab263784) | Abcam
Brand: Abcam Limited
SKU: AB263784
Price: 690 USD
Availability: InStock

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IRF3 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 deletion in exon5.

View Alternative Names

IIAE7, IRF3_HUMAN, Interferon regulatory factor 3, MGC94729

3 Images

Lab

Western blot - Human IRF3 knockout HeLa cell lysate (AB263784)

Lane 1 : Jurkat cell lysate (20 μg)

Lane 2 : MCF7 cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : IRF3 knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab68481 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab68481 was shown to react with IRF3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255345 (knockout cell lysate ab263784) was used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab68481 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IRF3 antibody [EPR2418Y] (<a href='/en-us/products/primary-antibodies/irf3-antibody-epr2418y-ab68481'>ab68481</a>) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 20 µg

Lane 2:

MCF7 cell lysate at 20 µg

Lane 2:

Western blot - Human IRF3 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-irf3-knockout-hela-cell-line-ab255345'>ab255345</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

IRF3 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 37 kDa,51 kDa

false

Lab

Western blot - Human IRF3 knockout HeLa cell lysate (AB263784)

Lane 1 : Jurkat cell lysate (20 μg)

Lane 2 : MCF7 cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : IRF3 knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab76409 was shown to react with IRF3 in wild-type HeLa. Loss of signal was observed when knockout cell line ab255345 (knockout cell lysate ab263784) was used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab76409 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IRF3 antibody [EP2419Y] (<a href='/en-us/products/primary-antibodies/irf3-antibody-ep2419y-ab76409'>ab76409</a>) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 20 µg

Lane 2:

MCF7 cell lysate at 20 µg

Lane 2:

Western blot - Human IRF3 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-irf3-knockout-hela-cell-line-ab255345'>ab255345</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

IRF3 knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Predicted band size: 47 kDa

Observed band size: 47 kDa,50 kDa

false

Sanger Sequencing - Human IRF3 knockout HeLa cell lysate (AB263784)

Sanger seq

Unknown

Sanger Sequencing - Human IRF3 knockout HeLa cell lysate (AB263784)

Allele-1 : 1 bp deletion in exon5

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 deletion in exon5.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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

IRF3

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.

IRF3 also known as Interferon Regulatory Factor 3 acts as an important transcription factor in the immune response. It has a molecular weight of approximately 47 kDa. The IRF3 protein is mainly expressed in the cytoplasm and nucleus of various cell types including immune cells such as macrophages and dendritic cells. The protein becomes activated through phosphorylation a process frequently identified in its phosphorylated form phospo-IRF3 or p-IRF3 which facilitates its role in immune function.

Biological function summary

IRF3 participates in the regulation of type I interferon (IFN) response a fundamental antiviral defense mechanism. IRF3 when phosphorylated forms a complex with CBP/p300 which then translocates to the nucleus to drive the expression of IFN-stimulated genes. This action strengthens the innate immune response and boosts the body's ability to counteract viral infections. Its activity and regulation are significant for maintaining a balanced immune response without excessive inflammation.

Pathways

IRF3 is involved in the Toll-like receptor (TLR) and RIG-I-like receptor (RLR) signaling pathways both essential in pathogen recognition and response. Within these pathways IRF3 interacts with proteins such as MAVS and TBK1 to propagate immune signaling. The activation of IRF3 in these pathways results in the production of type I interferons and other cytokines orchestrating an effective antiviral response. These interactions highlight the protein's central role in mediating immune signaling cascades.

IRF3's malfunction or deregulation can contribute to autoimmune diseases and antiviral deficiencies. Conditions such as systemic lupus erythematosus (SLE) and chronic hepatitis B infection are linked to IRF3 activity. During autoimmune responses or viral persistence the aberrant activation of IRF3 can lead to inappropriate immune responses. The connection with proteins like STAT1 in these conditions highlights the complex network IRF3 engages in facilitating its impact on disease progression and immune dysregulation.

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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

Visit the General protocols
Visit the Troubleshooting

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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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human IRF3 knockout HeLa cell lysate

Western blot - Human IRF3 knockout HeLa cell lysate (AB263784)

Western blot - Human IRF3 knockout HeLa cell lysate (AB263784)

Sanger Sequencing - Human IRF3 knockout HeLa cell lysate (AB263784)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Proteins & Peptides

Proteins & Peptides

Product promise