Human UBXN6 (UBXD1) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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UBXN6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
UBX domain containing 1, UBX domain containing 2, UBX domain protein 6, UBX domain-containing protein 1, UBX domain-containing protein 6, UBXD1, UBXDC2, UBXN6_HUMAN
- WB
Unknown
Western blot - Human UBXN6 (UBXD1) knockout HEK-293T cell line (AB266689)
Lanes 1-3 : Merged signal (red and green). Green - ab81555 observed at 50 kDa. Red - loading control ab181602 observed at 36 kDa.
ab81555 Anti-UBXN6/UBXD1 antibody [2F8-24] was shown to specifically react with UBXN6/UBXD1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266689 (knockout cell lysate ab258747) was used. Wild-type and UBXN6/UBXD1 knockout samples were subjected to SDS-PAGE. ab81555 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Anti-UBXN6/UBXD1 antibody [2F8-24] (<a href='/en-us/products/unavailable/ubxn6ubxd1-antibody-2f8-24-ab81555'>ab81555</a>) at 1/500 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
UBXN6 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human UBXN6 (UBXD1) knockout HEK-293T cell line (ab266689)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
false
- Sanger seq
Unknown
Sanger Sequencing - Human UBXN6 (UBXD1) knockout HEK-293T cell line (AB266689)
Homozygous : 1 bp insertion in exon 2
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
Zygosity
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
UBXN6 plays an important role in the regulation of protein turnover as it is involved in the ubiquitin-proteasome complex. It assists in the disassembly of protein substrates tagged for degradation contributing to the maintenance of protein quality control within the cell. Through interaction with p97/VCP UBXN6 helps in processes like endoplasmic reticulum-associated degradation (ERAD) important for cellular response to misfolded proteins.
Pathways
UBXN6's involvement extends to the autophagy and ubiquitin-proteasome pathways. In the ubiquitin-proteasome pathway it acts as a regulatory component favoring the removal of damaged or unnecessary proteins by acting with partners like p97/VCP. Through the autophagy pathway UBXN6 aids in cellular cleanup and recycling processes supporting cellular survival mechanisms during stress conditions. These pathways highlight UBXN6's integral role in protein quality control and cellular defense.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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