RAB3A KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Homozygote. 62 bp deletion.
Images
Key facts
- Cell type
- SK-N-FI
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Homozygote. 62 bp deletion
RAB3A KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Homozygote. 62 bp deletion.
Key facts
- Cell type
- SK-N-FI
- Species or organism
- Human
- Tissue
- Brain
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Homozygote. 62 bp deletion
- Disease
- Neuroblastoma
- Concentration
Properties
- Gene name
- RAB3A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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 + 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 duration
- A few days
- Appropriate short-term storage conditions
- -80°C
- Appropriate long-term storage conditions
- -196°C
Notes
Recommended control: Human wild-type SK-N-FI cell line ( ab288716). 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM + 10% FBS
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.
- 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x104cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
Rab3A also known as Ras-related protein Rab-3A acts as a small GTPase involved in regulating membrane trafficking. Its molecular mass is approximately 25 kDa. Rab3A is highly expressed in neurons particularly at synaptic vesicles. It cycles between an active GTP-bound state and an inactive GDP-bound state to regulate the docking and fusion of synaptic vesicles with the presynaptic membrane.
- Biological function summary
Rab3A facilitates neurotransmitter release by interacting with effector proteins such as RIM and Rabphilin-3A. It is part of a larger complex that includes the SNAREs which help mediate synaptic vesicle fusion. Through its regulatory action Rab3A ensures that neurotransmitters are released in a controlled manner during synaptic transmission which is essential for proper neuronal communication.
- Pathways
Rab3A participates in the synaptic vesicle cycle which is vital in neurotransmitter release. This cycle involves key proteins like Syntaxin and VAMP. Rab3A interacts with these SNARE proteins to participate in vesicle docking and priming. Additionally it plays a role in the calcium-dependent exocytosis pathway which relies on calcium ions triggering vesicle fusion at nerve terminals.
- Associated diseases and disorders
Rab3A associates with neurological conditions such as epilepsy and synaptic dysfunction. Dysfunction in Rab3A mechanisms can lead to improper neurotransmitter release and synaptic connectivity contributing to these conditions. The protein links to diseases through its interactions with proteins such as RIM which can also affect synaptic function and are implicated in certain neurodevelopmental disorders.
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Sanger Sequencing - Human RAB3A knockout SK-N-FI cell line (ab288708)
Homozygote. 62 bp deletion
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