Human AK2 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Unknown
Western blot - Human AK2 knockout HEK-293T cell line (AB266539)
Lanes 1 - 4 : Merged signal (red and green). Green - ab166901 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.ab166901 was shown to react with AK2 in wild-type HEK-293T cells in western blot with loss of signal observed in AK2 knockout cell line ab266539 (AK2 knockout cell lysate ab257825). Wild-type and AK2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab166901 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-AK2 antibody [EPR11388(B)] (<a href='/en-us/products/primary-antibodies/ak2-antibody-epr11388b-ab166901'>ab166901</a>) 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
Predicted band size: 26 kDa
Observed band size: 26 kDa
false
- WB
Lab
Western blot - Human AK2 knockout HEK-293T cell line (AB266539)
Lanes 1-4 : Merged signal (red and green). Green - ab157206 observed at 30 kDa. Red - loading control ab8245 observed at 36 kDa.
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 ab257825) was used. Wild-type and AK2 knockout samples were subjected to SDS-PAGE. ab157206 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-AK2 antibody [EPR11387(B)] (<a href='/en-us/products/primary-antibodies/ak2-antibody-epr11387b-ab157206'>ab157206</a>) 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
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: 26 kDa
Observed band size: 30 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human AK2 knockout HEK-293T cell line (AB266539)
Allele-1 : 8 bp deletion in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human AK2 knockout HEK-293T cell line (AB266539)
Allele-2 : 1 bp insertion in exon 1.
Complementary Products
Primary Antibodies
AB249383
Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free
RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)](https://content.abcam.com/products/images/ak2-antibody-epr11388b-bsa-and-azide-free-ab249383--immunocytochemistry-immunofluorescence-img77411.jpg)
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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, 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.
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
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.
Pathways
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.
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
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