Human TMUB1 partial knockout HeLa cell line
- Advanced Validation
- What is this?
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TMUB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2.
View Alternative Names
C7orf21, Chromosome 7 Open Reading Frame 21, DULP, Dendritic Cell-Derived Ubiquitin-Like Protein, HOPS, Hepatocyte odd protein shuttling protein, SB144, TMUB1_HUMAN, Transmembrane and ubiquitin-like domain-containing protein 1, UNQ763/PRO1555, Ubiquitin-like protein DULP, Ubiquitin-like protein SB144
- WB
Lab
Western blot - Human TMUB1 partial knockout HeLa cell line (AB265852)
Lanes 1- 3 : Merged signal (red and green). Green - ab180586 observed at 27 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab180586 was shown to react with TMUB1 in wild-type HeLa (Human epithelial line from cervix adenocarcinoma) cells in western blot. Loss of signal was observed when knockout cell line ab265852 (knockout cell lysate ab258237) was used. Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) and TMUB1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) 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. ab180586 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-TMUB1 antibody [EPR14066] (<a href='/en-us/products/primary-antibodies/tmub1-antibody-epr14066-ab180586'>ab180586</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Lane 2:
TMUB1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Lane 2:
Western blot - Human TMUB1 partial knockout HeLa cell line (ab265852)
Lane 3:
HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate cell lysate at 20 µg
Predicted band size: 26 kDa
Observed band size: 27 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TMUB1 partial knockout HeLa cell line (AB265852)
Allele-1 : 1 bp deletion in exon 2.
- Sanger seq
Lab
Sanger Sequencing - Human TMUB1 partial knockout HeLa cell line (AB265852)
Sequencing chromatogram displaying sequence edit in exon 2
- Sanger seq
Unknown
Sanger Sequencing - Human TMUB1 partial knockout HeLa cell line (AB265852)
Allele-2 : 1 bp insertion in exon 2.
Reactivity data
Product details
Knockout profile: Only the long form of the protein (lHOPS, 27 kDa) has been knocked-out from the parental cell line. The band observed in the KO lysate at 21 kDa is likely to represent a short form of the protein (sHOPS) (doi: 10.4161/cc.27054). We have not investigated the function of the remaining form of the protein.
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
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.
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
![Western blot - Anti-TMUB1 antibody [EPR14066] (AB180586)](https://content.abcam.com/products/images/tmub1-antibody-epr14066-ab180586--western-blot-img12696.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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