Human UBQLN1 (Ubiquilin) knockout HeLa cell line
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- WB
Supplier Data
Western blot - Human UBQLN1 (Ubiquilin) knockout HeLa cell line (AB266047)
Blocking and diluting buffer and concentration : 5% NFDM/TBST.
In Western blot, ab325320 was shown to bind specifically to UBQLN1. Target of interest was observed at 62 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in UBQLN1 knockout cell line (lane 2, knockout cell line ab266047 / knockout cell lysate ab259229).
The identity of the lower MW band at approximately 35 kDa is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes:
Western blot - Anti-Ubiquilin/UBQLN1 antibody [EPR29585-56] (<a href='/en-us/products/primary-antibodies/ubiquilin-ubqln1-antibody-epr29585-56-ab325320'>ab325320</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2:
Western blot - Human UBQLN1 (Ubiquilin) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-ubqln1-ubiquilin-knockout-hela-cell-lysate-ab259229'>ab259229</a>) at 20 µg
Lane 3:
786-O (human kidney epithelial cell) whole cell lysate at 20 µg
Lane 4:
HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution
Observed band size: 62 kDa,36 kDa
false
Exposure time: 37s
- Sanger seq
Unknown
Sanger Sequencing - Human UBQLN1 (Ubiquilin) knockout HeLa cell line (AB266047)
Allele-1 : 2 bp deletion in exon 1
- Sanger seq
Unknown
Sanger Sequencing - Human UBQLN1 (Ubiquilin) knockout HeLa cell line (AB266047)
Allele-2 : 1 bp insertion in exon 1.
Complementary Products
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
Quality control - STR analysis
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Ubiquilin/UBQLN1 serves as a shuttle factor a component in the intricate network regulating protein degradation. It associates with ubiquitinated proteins and guides them to proteasomes or autophagosomes for disposal. Moreover Ubiquilin forms a complex with heat shock proteins and proteasome components enhancing its role in managing cellular protein quality control. Its activity actively influences proteostasis ensuring the careful balance of protein synthesis and breakdown.
Pathways
Ubiquilin/UBQLN1 is integral to the ubiquitin-proteasome system and autophagy pathways. In these pathways Ubiquilin partners with proteins such as ubiquitin ligases and various adaptor proteins to mediate the clearance of misfolded or excess proteins therefore preventing cellular damage. Its interaction with the chaperone HSP70 is critical for delivering proteins tagged for degradation. Through these pathways Ubiquilin influences cellular stress responses and protein turnover maintaining cellular function and health.
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
Target data
Alternative Names
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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