Human IFNG knockout Jurkat cell line
- Advanced Validation
- What is this?
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(1 Publication)
IFNG KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 92bp deletion and 5bp insertion in exon 3 & intron 3-4.
View Alternative Names
IF 1, IFG, IFI, IFN immune, IFN-gamma, IFNG_HUMAN, Immune interferon, Interferon gamma, Type II Interferon
- WB
Supplier Data
Western blot - Human IFNG knockout Jurkat cell line (AB273746)
Lane 1 : Wild-type Jurkat Vehicle control : PMA (0 ng/mL, 6 h) and ionomycin (0 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 20 ug
Lane 2 : Wild-type Jurkat Treated : PMA (25 ng/mL, 6 h) and ionomycin (1 μg/mL, 6 h), BFA (5 g/ml, last 5 h) cell lysate, 20 ug
Lane 3 : IFNG knockout Jurkat Vehicle control : PMA (0 ng/mL, 6 h) and ionomycin (0 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 20 ug
Lane 4 : IFNG knockout Jurkat Treated : PMA (25 ng/mL, 6 h) and ionomycin (1 μg/mL, 6 h), BFA (5 μg/ml, last 5 h) cell lysate, 40 ug
Western blot : Anti-Interferon gamma antibody [EPR23991-53] staining at 1/1000 dilution; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab267369 was shown to bind specifically to Interferon gamma. A band was observed at 28 kDa in treated wild-type Jurkat cell lysates with no signal observed at this size in treated IFNG knockout cell line ab273746 (knockout cell lysate ab275521). To generate this image, wild-type and IFNG knockout Jurkat 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 HRP conjugated Goat anti-Rabbit (H+L) secondary antibody and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. Blot was developed with an ultra-high sensitivity ECL reagent.
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- NGS
Lab
Next Generation Sequencing - Human IFNG knockout Jurkat cell line (AB273746)
Allele-1 : 92bp deletion and 5bp insertion in exon 3 & intron 3-4
Reactivity data
Product details
Recommended control: Human wild-type Jurkat cell line (ab275468). 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.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 2x105 cells/mL is recommended.
- Do not allow cell density to exceed 3x106 cells/mL.
Culture medium
RPMI + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
This cytokine is significant in promoting macrophage activation enhancing the antigen presentation process and boosting the antimicrobial activity of phagocytes. IFN-γ is not part of a larger protein complex but works as a homodimer in signal transduction. Its production heightens the Th1 immune response by stimulating the differentiation of naïve T cells into Th1 cells which is essential for effective cellular immunity.
Pathways
IFN-γ is integrally involved in the JAK-STAT signaling pathway alongside another critical cytokine Interleukin-12. This pathway further amplifies the immune response by regulating the expression of genes associated with cellular defense mechanisms. IFN-γ also interacts with the NF-kB pathway influencing inflammation and the activation of further immune responses. These interactions show a network of cooperativity with proteins like STAT1 and NF-kB essential for executing its biological roles.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Nature communications 16:216 PubMed39746936
2025
Applications
Unspecified application
Species
Unspecified reactive species
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Product promise
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