JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB266293

Human VAMP8 (EDB) knockout HEK-293T cell line

Be the first to review this product! Submit a review

|

(0 Publication)

VAMP8 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 5 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
3 Images
Western blot - Human VAMP8 (EDB) knockout HEK-293T cell line (AB266293)
  • WB

Lab

Western blot - Human VAMP8 (EDB) knockout HEK-293T cell line (AB266293)

False colour image of Western blot : Anti-VAMP8/EDB antibody [EP2629Y] staining at 1/10000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76021 was shown to bind specifically to VAMP8/EDB. A band was observed at 11 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in VAMP8 knockout cell line ab266293 (knockout cell lysate ab257791). To generate this image, wild-type and VAMP8 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

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:

Western blot - Human VAMP8 (EDB) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-vamp8-edb-knockout-hek-293t-cell-lysate-ab257791'>ab257791</a>)

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

VAMP8 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human VAMP8 (EDB) knockout HEK-293T cell line (ab266293)

Predicted band size: 11 kDa

Observed band size: 11 kDa

false

Sanger Sequencing - Human VAMP8 (EDB) knockout HEK-293T cell line (AB266293)
  • Sanger seq

Unknown

Sanger Sequencing - Human VAMP8 (EDB) knockout HEK-293T cell line (AB266293)

Allele-1 : 5 bp deletion in exon 2

Sanger Sequencing - Human VAMP8 (EDB) knockout HEK-293T cell line (AB266293)
  • Sanger seq

Unknown

Sanger Sequencing - Human VAMP8 (EDB) knockout HEK-293T cell line (AB266293)

Allele-2 : 1 bp insertion in exon 2.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 5 bp deletion in exon 2

style
left-column

Complementary Products

Primary Antibodies

AB76021

Anti-VAMP8/EDB antibody [EP2629Y]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-VAMP8/EDB antibody [EP2629Y] (AB76021)

  • 5
  • 1
Cell Lines & Lysates

AB266260

Human SH3GL1 (EEN) knockout HEK-293T cell line

Advanced Validation

Western blot - Human SH3GL1 (EEN) knockout HEK-293T cell line (AB266260)

  • 0
  • 0
Cell Lines & Lysates

AB266300

Human CAMKK2 knockout HEK-293T cell line

Sanger Sequencing - Human CAMKK2 knockout HEK-293T cell line (AB266300)

  • 0
  • 0
Cell Lines & Lysates

AB266287

Human RRAGD (Rag D) knockout HEK-293T cell line

Advanced Validation

Western blot - Human RRAGD (Rag D) knockout HEK-293T cell line (AB266287)

  • 0
  • 0
style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

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.

style
left-column

What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab266293-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266293 Human VAMP8 (EDB) knockout HEK-293T cell line", "number":"AB266293-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab255449 Human wild-type HEK-293T cell line", "number":"AB266293-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab266293-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab266293 Human VAMP8 (EDB) knockout HEK-293T cell line", "number":"AB266293-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
VAMP8
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
style
left-column

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.

style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.
style
left-column
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column

Target data

See full target information Vesicle-associated membrane protein 8
style
left-column
style
left-column
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com