RAB5A KO cell line available to order. Free of charge wild type control provided.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
Alternative names
RAB 5, RAB5A member RAS oncogene family, RAB5A_HUMAN, RAS associated protein RAB5A, Ras-related protein Rab-5A
Recommended products
RAB5A KO cell line available to order. Free of charge wild type control provided.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- RAB5A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
- 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
Recommended control: Human wild-type A549 cell line (Human wild-type A549 cell line ab275463). 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:Hams F12 + 5% 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.
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x103-1x104 cells/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 6x104 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.
• Do not allow the cell density to exceed 7x104 cells/cm2.
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
Rab5 also known as Ras-related protein Rab-5A is a small GTPase of approximately 24 kDa. It is part of the Rab family of proteins and functions as an endosomal marker in cells. Rab5 is ubiquitously expressed but shows enriched expression in tissues with a high rate of endocytosis like the brain and liver. This protein plays a significant role in early endosome fusion and trafficking acting as an essential regulator of vesicular transport processes within the cell.
- Biological function summary
Rab5 regulates the early endocytic pathway by mediating vesicle docking and fusion processes. It is not a part of a large complex but interacts with a range of effector proteins that facilitate endosome maturation. Rab5's role as an endosomal marker makes it important for membrane trafficking receptor recycling and signal transduction. Researchers often identify Rab5 in cellular studies using endosome staining techniques or specific antibodies like Drosophila antibodies to visualize its function and distribution.
- Pathways
The endocytic network heavily involves Rab5 particularly influencing the endocytosis and endosomal signaling pathways. It frequently interacts with proteins such as EEA1 (Early Endosome Antigen 1) and Rabaptin-5 which are critical for endosomal membrane fusion. This mechanistic involvement ensures the correct transfer and processing of endocytic vesicles facilitating the continual cycle of membrane and receptor turnover in cells.
- Associated diseases and disorders
Rab5 dysfunction associates with neurological conditions such as Alzheimer's disease and cancer. Alterations in Rab5 levels or function can disrupt normal endosomal trafficking leading to amyloid beta peptide accumulation in Alzheimer's. Additionally Rab5's role in cancer relates to its regulation of growth factor receptors where its interaction with proteins like the PE marker influences tumor progression. Understanding Rab5 pathways and interactions paves the way for developing targeted therapeutics in these disorders.
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1 product image
Next Generation Sequencing - Human RAB5A knockout A549 cell line (ab301197)
62 bp insertion after Arg7 of the WT protein
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