VAMP8 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%.
Images
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%
Alternative names
ED B, Endobrevin, VAMP8_HUMAN, Vesicle-associated membrane protein 8
Recommended products
VAMP8 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%.
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%
- Disease
- Epidermoid Carcinoma
- Concentration
Properties
- Gene name
- VAMP8
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- 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 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
VAMP8 also known as synaptobrevin-like protein 2 or EDB is a SNARE protein with a mass of approximately 12 kDa. This protein is widely expressed in many tissues including the lung pancreas and kidney. It plays an important role in vesicle-mediated transport where it facilitates the fusion of vesicles with target membranes as part of a larger protein complex. By participating in these membrane fusion processes the protein contributes to cellular trafficking.
- Biological function summary
VAMP8 engages in vesicular transport mechanisms that support exocytosis and endocytosis processes. It forms part of the SNARE complex which is important for merging vesicles with their target compartments. Through this association VAMP8 aids the release of substances such as hormones and digestive enzymes from the cells. This process is critical for maintaining the balance and function of cellular activities across different tissues.
- Pathways
VAMP8 functions in the context of the trafficking pathways that include the endocytic and exocytic pathways. It is a participant in regulated exocytosis where it interacts with other SNARE proteins like syntaxin and SNAP-25 ensuring precise membrane fusion events occur. These pathways ensure proper cellular communication and debris clearance underlining the importance of VAMP8 in general cellular homeostasis.
- Associated diseases and disorders
VAMP8 is associated with conditions like inflammatory diseases and certain exocytic disorders. Researchers have observed its involvement in conditions like pancreatitis where its regulatory function in enzyme secretion becomes apparent. VAMP8 also has connections with other disease-related proteins such as those in the SNARE family which contribute to altered secretory pathways and inflammation. Understanding the precise mechanisms of VAMP8 helps in developing therapeutic approaches to managing these disorders.
Product promise
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2 product images
Western blot - Human VAMP8 (EDB) knockout A-431 cell line (ab269584)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-VAMP8/EDB antibody [EP2629Y] ab76021 observed at 13 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.Anti-VAMP8/EDB antibody [EP2629Y] ab76021 was shown to react with VAMP8/EDB in wild-type A431 cells in western blot. Loss of signal was observed when VAMP8 knockout sample was used. Wild-type and VAMP8 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-VAMP8/EDB antibody [EP2629Y] ab76021 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4° at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-VAMP8/EDB antibody [EP2629Y] (Anti-VAMP8/EDB antibody [EP2629Y] ab76021) at 1/10000 dilution
Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2: VAMP8 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human VAMP8 (EDB) knockout A-431 cell line (ab269584)
Lane 3: THP-1 (Human monocytic leukemia cell line) whole cell lysate at 20 µg
Lane 4: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 11 kDa
Observed band size: 13 kDa
Next Generation Sequencing - Human VAMP8 (EDB) knockout A-431 cell line (ab269584)
Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift = 99.7%
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