JAK2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and 5 bp deletion in exon 3.
JAK2_HUMAN, JTK 10, Janus Activating Kinase 2, Janus kinase 2, Janus kinase 2 (a protein tyrosine kinase), OTTHUMP00000043260, THCYT3, Tyrosine-protein kinase JAK2, kinase Jak2
JAK2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and 5 bp deletion in exon 3.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
JAK2 also known as Janus kinase 2 is a protein tyrosine kinase with a molecular weight of 125 kDa. It plays an essential mechanical role in the signaling pathways by acting as an intermediary between cell surface receptors and intracellular signaling molecules. The JAK2 protein binds to certain cytokine receptors facilitating signal transduction necessary for various cellular responses. JAK2 is expressed in many tissues including hematopoietic cells which are associated with the blood and immune systems.
JAK2 is important for transmitting signals that dictate cell growth survival and differentiation within the hematopoietic system. It operates bodily functions by forming complexes with specific phosphorylation sites on its associated receptors. Through this formation JAK2 influences the signaling cascade particularly by interacting with other signal transducers and activators where it phosphorylates and becomes activated. This action affects gene transcription directly correlated with cellular proliferation and differentiation processes.
The function of JAK2 integrates into the JAK-STAT signaling pathway which is an important pathway in the regulation of immune function growth and development. It works in conjunction with proteins such as STAT3 and STAT5 to transmit signals from cytokine receptors to the nucleus. This pathway critically impacts responses like inflammation and hematopoiesis aligning with its role in precursor proliferation within the bone marrow and various immune cells’ function.
JAK2 has significant implications in conditions like myeloproliferative neoplasms and polycythemia vera. Mutations in the JAK2 gene notably the JAK2 V617F mutation lead to uncontrolled cell division as they disrupt normal signaling mechanisms often resulting in blood cell disorders. In these contexts JAK2 interacts with proteins such as EpoR and MPL which play roles within these disease pathways. Understanding how JAK2 mutations contribute to disease progression offers pathways for targeted therapies.
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Terms & Conditions.
Anti-JAK2 antibody [EPR108(2)] ab108596 was shown to react with JAK2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267113 (knockout cell lysate Human JAK2 knockout A549 cell lysate ab256963) was used. Wild-type A549 and JAK2 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-JAK2 antibody [EPR108(2)] ab108596 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4° at a 1 in 1000 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-JAK2 antibody [EPR108(2)] (Anti-JAK2 antibody [EPR108(2)] ab108596) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: JAK2 knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human JAK2 knockout A549 cell line (ab267113)
Lane 3: K562 cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 131 kDa
Observed band size: 131 kDa
Allele-2: 1 bp insertion in exon 3.
Allele-1: 5 bp deletion in exon3
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