AK2 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 1 and 8 bp deletion in exon 1.
ADK2, ATP-AMP transphosphorylase, ATP-AMP transphosphorylase 2, Adenylate kinase 2, Adenylate kinase isoenzyme 2, EC 2.7.4.3, KAD2_HUMAN, adenylate kinase 2, mitochondrial, mitochondrial
AK2 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 1 and 8 bp deletion in exon 1.
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.
The adenylate kinase 2 also known as AK2 is an enzyme that catalyzes the conversion of adenine nucleotides within the cell. It facilitates the reversible conversion of ADP into ATP and AMP playing a critical role in cellular energy homeostasis. AK2 weighing approximately 26 kDa is primarily found in the mitochondrial intermembrane space of various tissues including liver muscle and brain. Its expression allows for efficient energy transfer essential for normal mitochondrial function and cellular metabolism.
AK2 functions in energy transfer and regulation in the cell serving as an important component in maintaining cellular ATP levels. AK2 is an integral part of the mitochondrial adenylate kinase system which manages energy flux in cells. By balancing the concentrations of ATP ADP and AMP AK2 directly influences cellular energy state and metabolic activity. The enzyme also supports nucleotide homeostasis therefore facilitating processes such as signal transduction and nucleic acid synthesis.
AK2 is involved in the adenylate kinase pathway important for energy metabolism. It coordinates with other enzymes like AK1 and AK3 ensuring efficient energy distribution in the cell. AK2 also interfaces with the apoptosis signaling pathways where it plays a part in regulating programmed cell death. The interaction of AK2 within these pathways emphasizes its essential role in maintaining cellular energy balance and undergoing apoptosis when necessary.
AK2 mutations link to rare conditions like reticular dysgenesis a severe form of combined immunodeficiency. The disorder arises when AK2 fails to initiate proper energy transfer necessary for white blood cell development. AK2 is also related to mitochondrial disorders where energy production gets critically impaired manifesting in muscle weakness and neurodegenerative symptoms. The enzyme's connection to proteins involved in energy pathways suggests its potential contribution to the progression of such diseases.
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All lanes: Western blot - Anti-AK2 antibody [EPR11388(B)] (Anti-AK2 antibody [EPR11388(B)] ab166901) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: AK2 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human AK2 knockout HEK-293T cell line (ab266539)
Lane 3: HL-60 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Anti-AK2 antibody [EPR11387(B)] ab157206 Anti-AK2 antibody [EPR11387(B)] was shown to specifically react with AK2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266539 (knockout cell lysate Human AK2 knockout HEK-293T cell lysate ab257825) was used. Wild-type and AK2 knockout samples were subjected to SDS-PAGE. Anti-AK2 antibody [EPR11387(B)] ab157206 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 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-AK2 antibody [EPR11387(B)] (Anti-AK2 antibody [EPR11387(B)] ab157206) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: AK2 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human AK2 knockout HEK-293T cell line (ab266539)
Lane 3: HL-60 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 26 kDa
Observed band size: 30 kDa
Allele-1: 8 bp deletion in exon 1
Allele-2: 1 bp insertion in exon 1.
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