Human DNAJC13 (RME-8) knockout HeLa cell line
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- Sanger seq
Unknown
Sanger Sequencing - Human DNAJC13 (RME-8) knockout HeLa cell line (AB265328)
Allele-1 : 2 bp deletion in exon 15.
- Sanger seq
Unknown
Sanger Sequencing - Human DNAJC13 (RME-8) knockout HeLa cell line (AB265328)
Allele-2 : 1 bp insertion in exon 15.
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
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 activity of RME-8 is essential for the maintenance of cellular homeostasis. It interacts with other proteins to form a complex that facilitates proper protein sorting within the endosomal network. RME-8's role in this intricate system ensures the appropriate recycling of receptors and disposal of damaged cellular components aiding in overall cell function. The protein regulates the spatial organization of endosomes an important process necessary for nutrient uptake and receptor recycling which impacts cell signaling and metabolism.
Pathways
RME-8 participates in endocytic pathways where it collaborates with proteins like Eps15 and Hsc70 to manage cargo trafficking. The endocytic pathway heavily relies on RME-8's functions for sustaining efficient internalization and routing of molecules such as receptors and ligands. Additionally the protein interacts with components of the retromer complex indicating a connection to the retromer-mediated transport pathway. This involvement may have effects on intracellular transport activities influencing the broader cellular communication network.
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'.
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