AK4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 2.
AK3L2, AK4, ATP-AMP transphosphorylase, Adenylate kinase 3 like 1, Adenylate kinase 3-like, Adenylate kinase isoenzyme 4, mitochondrial, GTP:AMP phosphotransferase, KAD4_HUMAN, MGC166959, adenylate kinase 4, mitochondrial adenylate kinase 3, nucleoside-triphosphate adenylate kinase
AK4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 2.
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.
AK3L1 also known as adenylate kinase 3-like 1 is an enzyme with a molecular mass of approximately 26 kDa. This protein primarily catalyzes the reversible transfer of phosphate groups between ATP and AMP. AK3L1 is expressed mainly in the mitochondrial matrix playing a fundamental role in nucleotide metabolism. By facilitating energy transfer within the mitochondria AK3L1 ensures the availability of ATP and maintains cellular energy homeostasis.
AK3L1 regulates mitochondrial energy metabolism and supports cellular functions. It operates as part of a network managing the mitochondrial adenylate kinase isoform family participating in mitochondrial phosphotransfer systems. AK3L1 contributes to sustaining energy balance especially in tissues with high-energy demand like muscle and brain. Within these systems AK3L1 interacts with other adenylate kinases and mitochondrial proteins to balance nucleotide ratios and prevent energy deficits.
AK3L1 integrates into the wider cellular energy regulation and oxidative phosphorylation pathways. It supports the cellular energy landscape by linking to the citric acid cycle and oxidative phosphorylation essential for ATP production. AK3L1 coordinates with proteins like AK2 influencing energy metabolism and ensuring effective cellular processes. By aligning with these pathways AK3L1 plays a significant role in ensuring efficient bioenergetic processes.
AK3L1 features involvement in conditions such as mitochondrial myopathy and neurodegenerative disorders. Its dysfunction can disrupt mitochondrial energy balance linking it to diseases characterized by energy production deficits. The protein closely interacts with AK2 and other mitochondrial enzymes whose imbalance can further contribute to disease development. Understanding AK3L1's role provides insight into potential therapeutic targets for these disorders addressing the underlying mitochondrial dysfunctions.
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Anti-AK3L1 antibody [EPR7678] ab131327 Anti-AK3L1 antibody [EPR7678] was shown to specifically react with Adenylate kinase 4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265331 (knockout cell lysate Human AK4 (AK3L1) knockout HeLa cell lysate ab257827) was used. Wild-type and Adenylate kinase 4 knockout samples were subjected to SDS-PAGE. Anti-AK3L1 antibody [EPR7678] ab131327 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° 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-AK3L1 antibody [EPR7678] (Anti-AK3L1 antibody [EPR7678] ab131327) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: AK3L1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human AK4 (AK3L1) knockout HeLa cell line (ab265331)
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: MCF7 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 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 25 kDa
Homozygous: 2 bp insertion in exon 2.
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