STAT2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2.
A549
Human
Lung
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2
Homo sapiens interferon alpha induced transcriptional activator, ISGF-3, MGC59816, P113, STAT113, STAT2_HUMAN, Signal transducer and activator of transcription 2, interferon alpha induced transcriptional activator, signal transducer and activator of transcription 2 113kD
STAT2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2.
A549
Human
Lung
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 2 and 1 bp deletion in exon 2
Puromycin 1µg/mL
Carcinoma
STAT2
Knockout
CRISPR technology
Sanger Sequencing, Western blot
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 1 US: 1
Adherent
Male
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.
F-12K + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type A549 cell line (Human wild-type A549 cell line ab255450). 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.
This supplementary information is collated from multiple sources and compiled automatically.
The Signal Transducer and Activator of Transcription 2 commonly known as STAT2 is a protein with a molecular mass of approximately 97 kDa. It functions mainly as a transcription factor and is an essential mediator of the interferon (IFN) signaling pathway. STAT2 is expressed in various tissues including immune cells where it responds to external stimuli. It works by translocating to the nucleus upon phosphorylation subsequently influencing the expression of genes involved in immune response.
Members of the STAT protein family including STAT2 play a central role in mediating immune responses. STAT2 often forms a complex with STAT1 and other proteins such as IRF9 to initiate the transcription of interferon-stimulated genes (ISGs). This complex known as the ISGF3 complex facilitates the cellular response to viral infections by promoting the expression of antiviral proteins that help modify the host cellular environment to resist viral replication.
The involvement of STAT2 is closely associated with the JAK-STAT signaling pathway and the interferon signaling pathway. In these pathways it interacts with proteins like JAK1 TYK2 and STAT1 playing a part in transmitting signals from the cell surface to the nucleus. Such pathways are fundamental to regulating the body's immune response to pathogens and managing cell growth and apoptosis in various cells.
STAT2 alterations have been linked to autoimmune diseases and viral infections. Disruptions in STAT2 function can impair the body's ability to combat viruses effectively making individuals more susceptible to infections. STAT2 mutations or dysregulations might also connect to disorders like Systemic Lupus Erythematosus where immune system malfunctions occur. The interactions of STAT2 with other proteins like STAT1 and IRF9 in disease contexts underline its importance in maintaining immune system balance.
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Lanes 1- 2: Merged signal (red and green). Green - Anti-STAT2 antibody [Y141] ab32367 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-STAT2 antibody [Y141] ab32367 was shown to react with STAT2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout sample Human STAT2 knockout A549 cell lysate ab257185 was used. Wild-type A549 and STAT2 knockout A549 cell lysates were subjected to SDS-PAGE. Anti-STAT2 antibody [Y141] ab32367 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4° at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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-STAT2 antibody [Y141] (Anti-STAT2 antibody [Y141] ab32367) at 1/5000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: STAT2 knockout A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 97 kDa
Observed band size: 97 kDa
Representative images of STAT2 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Allele-1: 19 bp deletion in exon2
Allele-2: 1 bp deletion in exon 2.
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