DNAJC13 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and 2 bp deletion in exon 15.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and 2 bp deletion in exon 15
Alternative names
DJC13_HUMAN, DNAJC13, DnaJ (Hsp40) homolog subfamily C member 13, DnaJ domain containing protein RME 8, DnaJ homolog subfamily C member 13, KIAA0678, RME-8, Required for receptor-mediated endocytosis 8
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DNAJC13 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and 2 bp deletion in exon 15.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and 2 bp deletion in exon 15
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- DNAJC13
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
RME-8 also known as DNAJC13 is a protein that functions as an ATPase assisting in the regulation of clathrin-mediated endocytosis. It possesses a mass of approximately 218 kDa and is mainly found in the central nervous system though it is also present in various other tissues. Mechanically RME-8 binds to clathrin-coated vesicles and plays a part in rearranging them during endocytosis. It interacts with multiple factors involved in membrane trafficking contributing to the modulation of vesicle fission and fusion events.
- 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.
- Associated diseases and disorders
RME-8 has connections to neurodegenerative disorders such as Parkinson's disease. Mutations in the DNAJC13 gene which codes for RME-8 have been linked to familial parkinsonism. The dysfunction of RME-8 affects endosomal processes potentially contributing to the pathophysiology of Parkinson's by impacting dopamine neuron survival. It is also implicated in Alzheimer's disease with disruptions in its role possibly affecting amyloid-beta processing through its interactions with proteins like Sortilin-related receptor 1 (SORL1) which is known to influence amyloid precursor protein sorting and degradation.
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2 product images
Sanger Sequencing - Human DNAJC13 (RME-8) knockout HeLa cell line (ab265328)
Allele-1: 2 bp deletion in exon 15.
Sanger Sequencing - Human DNAJC13 (RME-8) knockout HeLa cell line (ab265328)
Allele-2: 1 bp insertion in exon 15.
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