Human VAV3 knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Lanes 1 and 3:
Western blot - Anti-VAV3 antibody [EP1130Y] (<a href='/en-us/products/primary-antibodies/vav3-antibody-ep1130y-ab52938'>ab52938</a>) at 1/1000 dilution
Lane 2:
Western blot - Anti-VAV3 antibody [EP1130Y] (<a href='/en-us/products/primary-antibodies/vav3-antibody-ep1130y-ab52938'>ab52938</a>)
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
Predicted band size: 98 kDa
Observed band size: 98 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human VAV3 knockout HeLa cell line (AB265419)
Allele-1 : 1 bp deletion in exon 18.
- Sanger seq
Unknown
Sanger Sequencing - Human VAV3 knockout HeLa cell line (AB265419)
Allele-2 : 1 bp insertion in exon 18.
Complementary Products
![Flow Cytometry (Intracellular) - Anti-VAV3 antibody [EP1130Y] (AB52938)](https://content.abcam.com/products/images/vav3-antibody-ep1130y-ab52938--flow-cytometry-intracellular-img167047.jpg)
Primary Antibodies
AB315280
Anti-PDE3A antibody [EPR28228-6] - BSA and Azide free
Recombinant|RabMAb
![Western blot - Anti-PDE3A antibody [EPR28228-6] - BSA and Azide free (AB315280)](https://content.abcam.com/products/images/pde3a-antibody-epr28228-6-bsa-and-azide-free-ab315280--western-blot-img427398.jpg)
- 0
- 0
Primary Antibodies
AB1791
Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade
Lab Essentials
- 5
- 233
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, 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.
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
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.
Quality control
STR analysis
D5S818, TH01, D16S539, TPOX, CSF1PO, D13S317, D7S820
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
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