Human UFM1 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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UFM1 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: 23 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
BM 002, C13orf20, Chromosome 13 open reading frame 20, UFM1_HUMAN, Ubiquitin-fold modifier 1
- WB
Lab
Western blot - Human UFM1 knockout HEK-293T cell line (AB266782)
Western blot : Anti-UFM1 antibody [EPR4264(2)] (ab109305) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab109305 was shown to bind specifically to UFM1. A band was observed at 9/28 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in UFM1 knockout cell line. To generate this image, wild-type and UFM1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-UFM1 antibody [EPR4264(2)] (<a href='/en-us/products/primary-antibodies/ufm1-antibody-epr42642-ab109305'>ab109305</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
UFM1 knockout HEK-293T cell lysate at 20 µg
Lane 3:
U-2 OS cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 9 kDa,28 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human UFM1 knockout HEK-293T cell line (AB266782)
Homozygous : 23 bp insertion in exon2
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
UFM1 functions as a signaling molecule that influences various cellular pathways. It acts as part of a complex with other enzymes including UBA5 UFC1 and UFL1 which are important for its conjugation process. This modification system plays a role in endoplasmic reticulum (ER) stress response by affecting proteins involved in ER-associated degradation (ERAD). Its interaction with these proteins helps cells manage stress and maintain homeostasis.
Pathways
UFM1 participates in the ER stress response pathway and is also connected to the mTOR signaling pathway. Both pathways are essential for cell growth and survival. Within these pathways UFM1 interacts with proteins like mLST8 in the mTOR pathway integrating signals that coordinate cell metabolism growth and apoptosis. The precise regulation of these pathways highlights UFM1's role in 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
Target data
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UFM1 antibody [EPR4264(2)] (AB109305)](https://content.abcam.com/products/images/ufm1-antibody-epr42642-ab109305--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img91376.jpg)
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