Human MELK knockout HCT116 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human MELK knockout HCT116 cell line (AB266896)
Lanes 1 - 2 : Merged signal (red and green). Green - ab108529 observed at 75 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108529 was shown to react with MELK in wild-type HCT116 cells in western blot with loss of signal observed in MELK knockout cell line ab266896 (MELK knockout cell lysate ab257537). Wild-type and MELK knockout HCT116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108529 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-MELK antibody [EPR3981] (<a href='/en-us/products/primary-antibodies/melk-antibody-epr3981-ab108529'>ab108529</a>) at 1/1000 dilution
Lane 1:
HCT116 cell lysate at 20 µg
Lane 2:
MELK knockout HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human MELK knockout HCT116 cell line (ab266896)
Predicted band size: 74 kDa
Observed band size: 75 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MELK knockout HCT116 cell line (AB266896)
Allele-1 : 1 bp deletion in exon9
- Sanger seq
Unknown
Sanger Sequencing - Human MELK knockout HCT116 cell line (AB266896)
Allele-2 : Insertion of the selection cassette in exon 9.
Complementary Products
![Western blot - Anti-MELK antibody [EPR3981] (AB108529)](https://content.abcam.com/products/images/melk-antibody-epr3981-ab108529--western-blot-img1288.jpg)
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
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
McCoY5a + 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
MELK regulates signals that control the cell cycle apoptosis and embryonic development. MELK works as a component of complexes involved in maintaining cellular homeostasis. The protruding roles involve influencing cellular architecture and governance of stem cell maintenance. Studies in animal models show MELK involvement in maintaining pluripotency and regulation during the development phase.
Pathways
MELK participates in key processes such as the cell cycle and apoptosis pathways. Its diverse functionality connects MELK to proteins like Bcl-2-associated death promoter (BAD) and p53 which further influence main cellular events. These pathways and interactions make MELK a target of interest in the study of cellular growth and programmed cell death.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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