IFIT1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 25 bp deletion in exon 3.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 25 bp deletion in exon 3
Recommended products
IFIT1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 25 bp deletion in exon 3.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 25 bp deletion in exon 3
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- IFIT1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Viability
- ~ 80%
- Adherent/suspension
- Adherent
- Gender
- Male
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
All seeding densities should be based on cell counts gained by established methods.
Cells should be passaged when they have achieved 80-90% confluence (approx 8x104-1x105 cells/cm2).
- Culture medium
- F-12K + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (ab259777). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: F-12K + 10% FBS
Initial handling guidelines:
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10E4 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines:
• All seeding densities should be based on cell counts gained by established methods.
• A guide seeding density of 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
We will provide viable cells that proliferate on revival.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
IFIT1 also known as Interferon-Induced Protein with Tetratricopeptide Repeats 1 is a protein that plays a significant role in antiviral defense mechanisms. It has a molecular mass of approximately 56 kDa. Researchers find IFIT1 predominantly expressed in cells stimulated by interferons. The protein detects viral single-stranded RNA which in turn helps initiate the antiviral response. You will find IFIT1 in various cell types mainly those exposed to viral infections as it is an important part of the body's innate immune system.
- Biological function summary
Activities of IFIT1 focus on inhibiting viral replication and translation processes. The protein binds to viral RNAs via its distinctive sensor domains preventing infections from progressing. IFIT1 operates often as part of a larger protein complex with other IFIT family members improving its ability to intercept and inhibit viral RNA. This concerted action provides an important line of defense against diverse viral pathogens by interfering with viral protein synthesis.
- Pathways
IFIT1 occupies an important place in the interferon signaling pathways particularly the Jak-STAT pathway. This pathway is critical for mounting antiviral responses when viral infections occur. Through its interactions within the pathway IFIT1 works together with related proteins like IFIT2 and IFIT3 to repel viral threats. By integrating into these pathways it contributes to amplifying the immune response promoting a robust defense system against foreign viral invaders.
- Associated diseases and disorders
IFIT1 is associated mainly with viral diseases such as influenza and hepatitis C. Its role in these diseases involves preventing the communication and replication of the viruses making it a pivotal part of antiviral therapeutic research. Additionally proteins like RIG-I interact with IFIT1 during these processes to recognize viral RNA enhancing the effectiveness of the immune response. These protein partnerships drive ongoing research looking to leverage IFIT1's antiviral properties for treatment developments.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Western blot - Human IFIT1 knockout A549 cell line (ab266957)
Anti-IFIT1 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to IFIT1. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in IFIT1 knockout cell line ab266957 (knockout cell lysate ab258464). To generate this image, wild-type and IFIT1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Performed under reducing conditions.
Sanger Sequencing - Human IFIT1 knockout A549 cell line (ab266957)
Allele-1: 25 bp deletion in exon3
Sanger Sequencing - Human IFIT1 knockout A549 cell line (ab266957)
Allele-2: 10 bp deletion in exon 3.
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