Human UFC1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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UFC1 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: 16 bp deletion 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
CGI-126, HSPC155, UFC1_HUMAN, Ubiquitin-fold modifier-conjugating enzyme 1, Ufm1-conjugating enzyme 1
- WB
Lab
Western blot - Human UFC1 knockout HEK-293T cell line (AB266814)
Lanes 1- 2 : Merged signal (red and green). Green - ab189251 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab189251 was shown to react with UFC1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266814 (knockout cell lysate ab257781) was used. Wild-type HEK-293T and UFC1 HEK-293T KO 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. ab189251 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 10000 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-UFC1 antibody [EPR15014] (<a href='/en-us/products/primary-antibodies/ufc1-antibody-epr15014-ab189251'>ab189251</a>) at 1/10000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
UFC1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human UFC1 knockout HEK-293T cell line (ab266814)
Predicted band size: 19 kDa
Observed band size: 20 kDa
false
- WB
Lab
Western blot - Human UFC1 knockout HEK-293T cell line (AB266814)
Lanes 1- 2 : Merged signal (red and green). Green - ab189252 observed at 20 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab189252 was shown to react with UFC1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266814 (knockout cell lysate ab257781) was used. Wild-type HEK-293T and UFC1 HEK-293T KO 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. ab189252 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-UFC1 antibody [EPR15014-102] (<a href='/en-us/products/primary-antibodies/ufc1-antibody-epr15014-102-ab189252'>ab189252</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
UFC1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human UFC1 knockout HEK-293T cell line (ab266814)
Predicted band size: 19 kDa
Observed band size: 20 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human UFC1 knockout HEK-293T cell line (AB266814)
Homozygous : 16 bp deletion 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 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
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
UFC1 is involved in the process of UFMylation which modifies proteins through the attachment of UFM1 much like ubiquitination influences protein degradation and signaling. UFC1 forms part of a complex that also includes UBA5 and UFL1 the E1 activating enzyme and E3 ligase respectively. This UFMylation system is necessary for endoplasmic reticulum-associated degradation (ERAD) contributing to the maintenance of protein homeostasis within cells. The disruption of this modification process can impact cellular stress response and protein quality control mechanisms.
Pathways
UFC1 plays a critical role in the ER stress response and protein quality control pathways. Within these pathways UFC1 interacts with proteins such as UFM1 and UFL1 to mediate the UFMylation of target proteins during cellular stress. Additionally UFC1 has been linked to pathways regulating autophagy where it indirectly influences the degradation of defective proteins. This interaction ultimately affects the cellular response to misfolded proteins and is pivotal for maintaining cellular equilibrium.
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
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