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AB270707

Human VAMP8 (EDB) knockout A-431 cell lysate

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VAMP8 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%.

View Alternative Names

ED B, Endobrevin, VAMP8_HUMAN, Vesicle-associated membrane protein 8

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Western blot - Human VAMP8 (EDB) knockout A-431 cell lysate (AB270707)
  • WB

Lab

Western blot - Human VAMP8 (EDB) knockout A-431 cell lysate (AB270707)

Lane 1 : Wild-type A431 cell lysate 20 ug

Lane 2 : VAMP8 knockout A431 cell lysate 20 ug

Lane 3 : THP-1 cell lysate 20 ug

Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate 20 ug

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 A-431 cells in western blot Loss of signal was observed when VAMP8 knockout cell line ab269584 (knockout cell lysate ab270707) was used. Wild-type and VAMP8 knockout A-431 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°C 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 (<a href='/en-us/products/cell-lines/human-vamp8-edb-knockout-a-431-cell-line-ab269584'>ab269584</a>)

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

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Next Generation Sequencing - Human VAMP8 (EDB) knockout A-431 cell lysate (AB270707)
  • NGS

Supplier Data

Next Generation Sequencing - Human VAMP8 (EDB) knockout A-431 cell lysate (AB270707)

Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift = 99.7%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift = 99.7%

Disease

Epidermoid Carcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
VAMP8
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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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

AB269584

Human VAMP8 (EDB) knockout A-431 cell line

Advanced Validation

Western blot - Human VAMP8 (EDB) knockout A-431 cell line (AB269584)

  • 0
  • 0
Primary Antibodies

AB181151

Anti-SNAP29 antibody [EPR9198(2)]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-SNAP29 antibody [EPR9198(2)] (AB181151)

  • 5
  • 1
Cell Lines & Lysates

AB263930

Human PICALM knockout U-2 OS cell lysate

Western blot - Human PICALM knockout U-2 OS cell lysate (AB263930)

  • 0
  • 0

Product promise

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