Human UBE2K (HIP2/LIG) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human UBE2K (HIP2/LIG) knockout HeLa cell line (AB266031)
Lanes 1-4 : Merged signal (red and green). Green - ab52930 observed at 26 kDa. Red - loading control ab8245 observed at 36 kDa.
ab52930 Anti-HIP2/LIG antibody [EP1145Y] was shown to specifically react with HIP2/LIG in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266031 (knockout cell lysate ab257778) was used. Wild-type and HIP2/LIG knockout samples were subjected to SDS-PAGE. ab52930 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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-HIP2/LIG antibody [EP1145Y] (<a href='/en-us/products/primary-antibodies/hip2-lig-antibody-ep1145y-ab52930'>ab52930</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
UBE2K knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human UBE2K (HIP2/LIG) knockout HeLa cell line (ab266031)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 22 kDa
Observed band size: 26 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human UBE2K (HIP2/LIG) knockout HeLa cell line (AB266031)
Homozygous : 1 bp deletion in exon1
Complementary Products
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
The function of HIP2 extends to protein homeostasis essential for maintaining cellular integrity. As part of the ubiquitin-proteasome system it ensures the elimination of damaged or excess proteins effectively preventing cellular stress or dysfunction. HIP2 operates within a complex that includes E1 enzymes and various E3 ligases highlighting its collaborative nature in protein turnover. The protein activity helps regulate cell cycle progression DNA repair and signal transduction ensuring normal cell function and adaptation to environmental changes.
Pathways
HIP2 is most involved in the ubiquitin-proteasome pathway and is linked to protein catabolism. It collaborates with key proteins like UBE2C and UBE2D which participate in tagging proteins for degradation. The coordination with these proteins demonstrates HIP2's role in managing protein levels in cells having implications in the cell's response to damage and stress. Additionally HIP2 intersects with pathways associated with cell cycle regulation allowing cells to maintain orderly division and prevent the accumulation of aberrant proteins.
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
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