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VASP KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2.

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Images

Western blot - Human VASP knockout HeLa cell line (AB265892), expandable thumbnail
  • Cell Culture - Human VASP knockout HeLa cell line (AB265892), expandable thumbnail
  • Sanger Sequencing - Human VASP knockout HeLa cell line (AB265892), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2

Alternative names

Recommended products

VASP KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2.

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2
Antibiotic resistance
Puromycin 1µg/mL
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
VASP
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

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

Cell culture

Biosafety level
EU: 2 US: 2
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

Recommended control: Human wild-type HeLa cell line (ab255928). 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.

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

VASP also known as Vasodilator-Stimulated Phosphoprotein is a 46-50 kDa protein involved in actin filament assembly and elongation. It plays an important role in cellular motility and the maintenance of cell shape. VASP is expressed in many cell types but is notably present in platelets and endothelial cells. It is an important player in signal transduction pathways that lead to cytoskeletal reorganization influencing cell movement and adhesion dynamics.

Biological function summary

VASP is an important component in cell movement regulation. It acts within a larger complex of actin-binding proteins that control the assembly of actin filaments. This protein influences cell adhesion migration and signal transduction processes. Through these actions VASP influences cellular responses to environmental cues making it essential for cellular dynamics and integrity during development and wound healing.

Pathways

VASP connects to significant signaling pathways such as the PI3K/Akt pathway and the cAMP pathway. It often interacts with proteins like c-Abl tyrosine kinase and Mena contributing to actin dynamics regulation. These pathways are critical for transducing signals from extracellular environments to intracellular responses allowing for coordinated cellular functions in processes such as cell migration and growth.

Associated diseases and disorders

VASP plays a role in cardiovascular disease and cancer-related metastasis due to its function in cell motility and adhesion. Abnormal VASP activity is associated with impaired endothelial cell function and contributes to atherosclerosis development. Additionally it interacts with proteins like Mena and fascin in cancer cells aiding in metastatic progression by enhancing motility and invasive capabilities.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Western blot - Human VASP knockout HeLa cell line (ab265892), expandable thumbnail

    Western blot - Human VASP knockout HeLa cell line (ab265892)

    Lanes 1-4: Merged signal (red and green). Green - Anti-VASP antibody [EPR1337(2)] ab109321 observed at 46 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    Anti-VASP antibody [EPR1337(2)] ab109321 Anti-VASP antibody [EPR1337(2)] was shown to specifically react with VASP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265892 (knockout cell lysate Human VASP knockout HeLa cell lysate ab257792) was used. Wild-type and VASP knockout samples were subjected to SDS-PAGE. Anti-VASP antibody [EPR1337(2)] ab109321 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-VASP antibody [EPR1337(2)] (Anti-VASP antibody [EPR1337(2)] ab109321) at 1/1000 dilution

    Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 2: VASP knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 2: Western blot - Human VASP knockout HeLa cell line (ab265892)

    Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

    Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Predicted band size: 39 kDa

    Observed band size: 46 kDa

  • Cell Culture - Human VASP knockout HeLa cell line (ab265892), expandable thumbnail

    Cell Culture - Human VASP knockout HeLa cell line (ab265892)

    Representative images of VASP knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

  • Sanger Sequencing - Human VASP knockout HeLa cell line (ab265892), expandable thumbnail

    Sanger Sequencing - Human VASP knockout HeLa cell line (ab265892)

    Homozygous: 13 bp deletion in exon 2.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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

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