Human WDR45 knockout HeLa cell line
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WDR45 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 3 and 1 bp insertion in exon 3.
View Alternative Names
JM5, NBIA4, WD repeat containing protein 45, WD repeat domain 45, WD repeat domain phosphoinositide interacting protein 4, WD repeat domain, X linked 1, WD45 repeat protein interacting with phosphoinositides 4, WDRX1, WDRXI4
- Sanger seq
Unknown
Sanger Sequencing - Human WDR45 knockout HeLa cell line (AB265981)
Allele-1 : 11 bp deletion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human WDR45 knockout HeLa cell line (AB265981)
Allele-2 : 1 bp insertion in exon 3.
Product details
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
WDR45 contributes significantly to the regulation of autophagy. It acts as part of a multi-protein complex referred to as the autophagy-related 2 (ATG) complex. This complex is involved in autophagosome formation which is an important component in cellular degradation pathways. WDR45 binds phosphoinositides which are phospholipids integral to vesicle trafficking and membrane dynamics during autophagy.
Pathways
The influence of WDR45 is evident in the PI3K/Akt signaling and AMPK signaling pathways. These pathways regulate cellular energy homeostasis and the response to metabolic stress. PI3K/Akt and AMPK modulate the activity of the mTOR pathway which is known to suppress autophagy. WDR45 interacts with other autophagy-related proteins such as LC3 and ULK1 facilitating the initiation of the autophagic process.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
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