Human ATOX1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ATOX1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 1 and 5 bp deletion in exon 1. 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
ATOX1_HUMAN, ATX1 antioxidant protein 1 homolog, ATX1 antioxidant protein 1 homolog (yeast), Copper transport protein, Copper transport protein ATOX1, HAH1, MGC138453, MGC138455, Metal transport protein, Metal transport protein ATX1
- WB
Lab
Western blot - Human ATOX1 knockout HEK-293T cell line (AB266651)
Lanes 1-4 : Merged signal (red and green). Green - ab154179 observed at 7 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154179 Anti-ATOX1 antibody [EPR10352] was shown to specifically react with ATOX1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266651 (knockout cell lysate ab257849) was used. Wild-type and ATOX1 knockout samples were subjected to SDS-PAGE. ab154179 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-ATOX1 antibody [EPR10352] (<a href='/en-us/products/primary-antibodies/atox1-antibody-epr10352-ab154179'>ab154179</a>) at 1/500 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ATOX1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ATOX1 knockout HEK-293T cell line (ab266651)
Lane 3:
HeLa 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: 36 kDa,45 kDa,62 kDa,7 kDa
Observed band size: 48 kDa,50 kDa,7 kDa,70 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATOX1 knockout HEK-293T cell line (AB266651)
Allele-1 : 11 bp deletion in exon 1
- Cell Culture
Unknown
Cell Culture - Human ATOX1 knockout HEK-293T cell line (AB266651)
Representative images of ATOX1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ATOX1 knockout HEK-293T cell line (AB266651)
Allele-2 : 5 bp deletion in exon 1.
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
ATOX1 serves as a chaperone that transports copper ions to ATPase proteins like ATP7A and ATP7B in the secretory pathway. ATOX1 works individually rather than as part of a larger protein complex handling copper metabolism effectively. By facilitating the delivery of copper to essential enzymes ATOX1 assists in maintaining critical cellular functions such as oxidative stress response and energy production.
Pathways
Copper ion transport involving ATOX1 plays an important role in the regulation of metal ion homeostasis pathways. ATOX1 interacts closely with ATP7A and ATP7B transporting copper ions necessary for redox reactions and enzyme activities in these pathways. ATOX1's role in copper regulation also aligns with antioxidant defense processes helping protect cells from damage due to reactive oxygen species.
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
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATOX1 antibody [EPR10352] (AB154179)](https://content.abcam.com/products/images/atox1-antibody-epr10352-ab154179--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img133389.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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