UFM1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp insertion in exon 2.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp insertion in exon 2
Alternative names
BM 002, C13orf20, Chromosome 13 open reading frame 20, UFM1_HUMAN, Ubiquitin-fold modifier 1
Recommended products
UFM1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp insertion in exon 2.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 23 bp insertion in exon 2
- Concentration
Properties
- Gene name
- UFM1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
UFM1 also called ubiquitin-fold modifier 1 is a small protein modifier with a mass of approximately 9 kDa. It is part of the ubiquitin-like protein family and primarily expressed in a wide range of tissues. UFM1 is involved in a unique form of post-translational modification known as UFMylation where it conjugates to substrate proteins altering their function and stability. This process plays a role in regulating cell cycle and apoptosis among other cellular processes.
- 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.
- Associated diseases and disorders
UFM1 has been linked to cancer and congenital disorders. Altered UFMylation due to mutations in related genes can contribute to tumor development by affecting protein degradation and cellular stress responses. In hereditary conditions dysfunctional UFM1 activity can lead to rare disorders affecting metabolism and neurological function. Proteins like mLST8 which interact with UFM1 in pathways can also play a role in these disease mechanisms suggesting potential targets for therapeutic intervention.
Product promise
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2 product images
Western blot - Human UFM1 knockout HEK-293T cell line (ab266782)
Western blot: Anti-UFM1 antibody [EPR4264(2)] (Anti-UFM1 antibody [EPR4264(2)] ab109305) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-UFM1 antibody [EPR4264(2)] 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)] (Anti-UFM1 antibody [EPR4264(2)] ab109305) 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
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 9 kDa, 28 kDa
Sanger Sequencing - Human UFM1 knockout HEK-293T cell line (ab266782)
Homozygous: 23 bp insertion in exon2
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