IRS2 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9.
IRS2_HUMAN, Insulin receptor substrate 2
IRS2 KO cell line available to order. KO validated by Immunocytochemistry, Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9.
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.
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.
Insulin Receptor Substrate 2 (IRS2) is an important cytoplasmic adaptor protein involved in insulin signaling. Alternate names for IRS2 include IRS-2 or HEK293 products IRS2. It plays a role in transducing signals from the insulin receptor tyrosine kinase to downstream effectors. IRS2 has a molecular mass of approximately 180 kDa. Expression of IRS2 occurs in many tissues with significant presence in liver and skeletal muscle indicating its broad physiological role.
IRS2 acts in signal transduction and is a part of the insulin and insulin-like growth factor signaling pathways. IRS2 does not function alone but interacts with other molecules to propagate signals that regulate cellular processes. It is essential for glucose homeostasis and lipid metabolism playing a critical role in maintaining energy balance within the organism. IRS2 also mediates other biological processes like cell growth and differentiation.
With regards to pathways IRS2 is involved in the insulin signaling and mTOR pathways. It serves as an important junction between metabolic and growth-promoting signals. Activation of IRS2 through insulin receptors leads to downstream signaling involving PI3K and Akt essential for mediating metabolic actions of insulin. It also interacts with mTOR a central protein in cell growth regulation. These pathways illustrate the integration of IRS2 in broader cellular responses to growth signals and energy availability.
IRS2 is linked to conditions like type 2 diabetes and obesity. Dysregulation of IRS2-mediated signaling can contribute to insulin resistance a characteristic feature of type 2 diabetes. In such disorders IRS2 signaling may be impaired or altered affecting its interaction with proteins like PI3K and Akt which are also implicated in these conditions. Furthermore chronic imbalance in IRS2 activity may contribute to obesity as it affects key metabolic pathways that regulate fat storage and energy expenditure.
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Terms & Conditions.
Anti-IRS2 antibody [EPR904(2)] ab134101 was shown to react with IRS2 in wild-type HEK-293 cells in western blot with loss of signal observed in IRS2 knockout sample. Wild-type and IRS2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-IRS2 antibody [EPR904(2)] ab134101 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-IRS2 antibody [EPR904(2)] (Anti-IRS2 antibody [EPR904(2)] ab134101) at 1/1000 dilution
Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2: IRS2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2: Western blot - Human IRS2 knockout HEK-293 cell line (ab264013)
Lane 3: A-375 cell lysate at 20 µg
Lane 4: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 137 kDa, 25 kDa
Observed band size: 160 kDa
Anti-IRS2 antibody [EPR1650(2)] ab133276 was shown to react with IRS2 in wild-type HEK-293 cells in western blot with loss of signal observed in IRS2 knockout sample. Wild-type and IRS2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-IRS2 antibody [EPR1650(2)] ab133276 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-IRS2 antibody [EPR1650(2)] (Anti-IRS2 antibody [EPR1650(2)] ab133276) at 1/1000 dilution
Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2: IRS2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2: Western blot - Human IRS2 knockout HEK-293 cell line (ab264013)
Lane 3: A-375 cell lysate at 20 µg
Lane 4: A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 160 kDa
Anti-IRS2 antibody [EPR904(2)] ab134101 staining IRS2 in wild-type HEK-293 cells (top panel) and IRS2 knockout HEK-293 cells (ab264013) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-IRS2 antibody [EPR904(2)] ab134101 at 1/500 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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