Human VAMP8 (EDB) knockout A-431 cell line
- Advanced Validation
- What is this?
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VAMP8 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
ED B, Endobrevin, VAMP8_HUMAN, Vesicle-associated membrane protein 8
- WB
Lab
Western blot - Human VAMP8 (EDB) knockout A-431 cell line (AB269584)
Lanes 1 - 4 : Merged signal (red and green). Green - ab76021 observed at 13 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
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 ab76021 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/vamp8-edb-antibody-ep2629y-ab76021'>ab76021</a>) 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
Predicted band size: 11 kDa
Observed band size: 13 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human VAMP8 (EDB) knockout A-431 cell line (AB269584)
Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift = 99.7%
Reactivity data
Product details
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.
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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![Flow Cytometry (Intracellular) - Anti-VAMP8/EDB antibody [EP2629Y] (AB76021)](https://content.abcam.com/products/images/vamp8-edb-antibody-ep2629y-ab76021--flow-cytometry-intracellular-img13849.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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