Human ADK knockout HeLa cell line
- Advanced Validation
- What is this?
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ADK KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 2 and 41 bp deletion in exon 2.
View Alternative Names
2310026J05Rik, 5033405D03Rik, ADK_HUMAN, AI255373, AI987814, AK, Adenosine 5''-phosphotransferase, Adenosine kinase, MGC6593, OTTHUMP00000019864, OTTHUMP00000019865
- WB
Lab
Western blot - Human ADK knockout HeLa cell line (AB265646)
Lanes 1- 2 : Merged signal (red and green). Green - ab243636 observed at 41 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab243636 was shown to react with Adenosine kinase in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265646 (knockout cell lysate ab257343) was used. Wild-type HeLa and ADK knockout HeLa 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. ab243636 and 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 (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-ADK antibody [EPR23166-143] (<a href='/en-us/products/primary-antibodies/adk-antibody-epr23166-143-ab243636'>ab243636</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ADK knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ADK knockout HeLa cell line (ab265646)
Predicted band size: 41 kDa
Observed band size: 41 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ADK knockout HeLa cell line (AB265646)
Allele-2 : 13 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human ADK knockout HeLa cell line (AB265646)
Allele-1 : 41 bp deletion in exon 2.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
The enzyme modulates cellular adenosine levels which influences various physiological functions such as sleep regulation and immune responses. ADK does not operate as part of a larger protein complex but acts independently to maintain intracellular energy balance by regulating the pools of adenosine and AMP. The enzyme activity affects purine metabolism important in diverse cellular activities.
Pathways
The activity of adenosine kinase fits prominently into the purine metabolism and energy homeostasis pathways. Its function maintains the cellular concentrations of adenosine a nucleotide affecting cellular signaling pathways including the AMP-activated protein kinase (AMPK) pathway. ADK interacts with enzymes like adenosine deaminase which collectively orchestrate the purine salvage pathway—pivotal in nucleotide recycling.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB243636
Anti-ADK antibody [EPR23166-143]
BOND RX™ Validated|RabMAb|Recombinant|KO Validated|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADK antibody [EPR23166-143] (AB243636)](https://content.abcam.com/products/images/adk-antibody-epr23166-143-ab243636--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img13990.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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