VAV3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 18 and 1 bp insertion in exon 18.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 18 and 1 bp insertion in exon 18
Alternative names
FLJ40431, Guanine nucleotide exchange factor VAV3, Protein vav 3, RGD1565941, VAV 3 oncogene, VAV 3 protein, VAV3_HUMAN, Vav 3 guanine nucleotide exchange factor
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VAV3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 18 and 1 bp insertion in exon 18.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 18 and 1 bp insertion in exon 18
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- VAV3
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- D5S818, TH01, D16S539, TPOX, CSF1PO, D13S317, D7S820
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
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The VAV3 protein also known as the guanine nucleotide exchange factor VAV3 functions as a regulator of Rho family GTPases by promoting the exchange of GDP for GTP. It weighs approximately 101 kDa and exhibits expression in various tissues including the brain heart and hematopoietic cells. VAV3 serves as an important modulator within the intracellular signaling processes by influencing actin cytoskeletal dynamics and impacting cellular morphology and movement.
- Biological function summary
VAV3 influences multiple cellular processes like cell proliferation differentiation and migration. Part of many signaling complexes VAV3 interacts with receptors on the cell surface transmitting signals inside the cell. These interactions play a role in the regulation of immune responses and its involvement in the nervous system aids in neuron development and connectivity.
- Pathways
VAV3 takes part in the regulation of the Rho GTPase pathway and the phosphoinositide 3-kinase (PI3K) pathway. VAV3 cooperates with proteins such as RAC1 and CDC42 influencing these pathways to modulate cell survival cytoskeleton rearrangement and membrane trafficking. Interaction with PI3K pathway components amplifies signals for growth factor-induced cellular responses impacting the overall cell function and homeostasis.
- Associated diseases and disorders
VAV3 influences both cancer and autoimmune diseases due to its regulatory role in cell signaling. Overexpression or abnormal regulation of VAV3 relates to prostate cancer progression and metastasis through its interaction with androgen receptor signaling pathways. In autoimmune diseases VAV3's interaction with proteins such as SYK can affect the severity of conditions like rheumatoid arthritis by modulating immune cell activation and response.
Product promise
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3 product images
Western blot - Human VAV3 knockout HeLa cell line (ab265419)
False colour image of Western blot: Anti-VAV3 antibody [EP1130Y] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-VAV3 antibody [EP1130Y] ab52938 was shown to bind specifically to VAV3. A band was observed at 98 kDa in wild-type HeLa cell lysates with no signal observed at this size in VAV3 knockout cell line ab265419 (knockout cell lysate Human VAV3 knockout HeLa cell lysate ab258280). To generate this image wild-type and VAV3 knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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 (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) at 1/20000 dilution.
Lanes 1 and 3: Western blot - Anti-VAV3 antibody [EP1130Y] (Anti-VAV3 antibody [EP1130Y] ab52938) at 1/1000 dilution
Lane 2: Western blot - Anti-VAV3 antibody [EP1130Y] (Anti-VAV3 antibody [EP1130Y] ab52938)
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Empty
Lane 2: Western blot - Human VAV3 knockout HeLa cell line (ab265419)
Lane 3: VAV3 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 98 kDa
Sanger Sequencing - Human VAV3 knockout HeLa cell line (ab265419)
Allele-1: 1 bp deletion in exon 18.
Sanger Sequencing - Human VAV3 knockout HeLa cell line (ab265419)
Allele-2: 1 bp insertion in exon 18.
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