Human AK1 knockout HeLa cell line
- Advanced Validation
- What is this?
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AK1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 4 and 1 bp deletion in exon 4 and 5 bp deletion in exon 4. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
AK 1, ATP-AMP transphosphorylase, ATP-AMP transphosphorylase 1, Adenylate kinase 1, Adenylate kinase isoenzyme 1, Adenylate kinase soluble, KAD1_HUMAN, Myokinase
- WB
Unknown
Western blot - Human AK1 knockout HeLa cell line (AB265132)
Lanes 1-3 : Merged signal (red and green). Green - ab138504 observed at 25 kDa. Red - loading control ab8245 observed at 36 kDa.
ab138504 Anti-AK 1 antibody [EP7083] was shown to specifically react with AK 1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265132 (knockout cell lysate ab257824) was used. Wild-type and AK 1 knockout samples were subjected to SDS-PAGE. ab138504 and 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 (ab216773) and 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-AK 1 antibody [EP7083] (<a href='/en-us/products/primary-antibodies/ak-1-antibody-ep7083-ab138504'>ab138504</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
AK1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human AK1 knockout HeLa cell line (ab265132)
Lane 3:
U-87MG cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 22 kDa
Observed band size: 25 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human AK1 knockout HeLa cell line (AB265132)
Allele-3 : 1 bp deletion in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human AK1 knockout HeLa cell line (AB265132)
Allele-1 : 14 bp deletion in exon 4.
- Sanger seq
Unknown
Sanger Sequencing - Human AK1 knockout HeLa cell line (AB265132)
Allele-2 : 5 bp deletion in exon 4.
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Adenylate kinase 1 is essential for maintaining the adenine nucleotide pool balance. It does not act alone but functions within diverse cellular contexts ensuring efficient energy supply which is necessary for critical cellular processes. In these contexts AK1 forms part of a larger network that involves other kinases and molecules operating to sustain sufficient energy levels during periods of high activity or stress. This orchestration of molecules ensures cells adapt to changing energy requirements swiftly.
Pathways
AK1 is a critical component of the ATP-generating phosphotransfer network and is involved in the cellular adenylate kinase reaction pathway. It interacts closely with proteins such as AMP-activated protein kinase (AMPK) which assists in detecting and responding to energy deficits. In the context of the phosphotransfer network maintaining the balance between ATP ADP and AMP is important and AK1 enables this dynamic equilibrium. By interrelating with these pathways AK1 supports efficient energy transduction and cellular adaptation to metabolic changes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Western blot - Anti-AK 1 antibody [EP7083] (AB138504)](https://content.abcam.com/products/images/ak-1-antibody-ep7083-ab138504--western-blot-img146093.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com