Human GAS6 knockout HEK-293 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human GAS6 knockout HEK-293 cell line (AB275837)
False colour image of Western blot : Anti-GAS 6 antibody [EPR23931-27] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab264098 was shown to bind specifically to GAS 6. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in GAS6 knockout cell line ab275837 (knockout cell lysate ab275811). To generate this image, wild-type and GAS6 knockout HEK-293 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-GAS 6 antibody [EPR23931-27] (<a href='/en-us/products/primary-antibodies/gas-6-antibody-epr23931-27-ab264098'>ab264098</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293 cell lysate at 20 µg
Lane 2:
GAS6 knockout HEK-293 cell lysate at 20 µg
Lane 2:
Western blot - Human GAS6 knockout HEK-293 cell line (ab275837)
Lane 3:
THP-1 cell lysate at 20 µg
Predicted band size: 80 kDa
Observed band size: 75 kDa
false
- NGS
Lab
Next Generation Sequencing - Human GAS6 knockout HEK-293 cell line (AB275837)
1 bp insertion after Glu162 of the WT protein
Complementary Products
Primary Antibodies
AB264098
Anti-GAS 6 antibody [EPR23931-27]
20ul selling size|RabMAb|Recombinant|KO Validated|Trial Size
![Flow Cytometry (Intracellular) - Anti-GAS 6 antibody [EPR23931-27] (AB264098)](https://content.abcam.com/products/images/gas-6-antibody-epr23931-27-ab264098--flow-cytometry-intracellular-img131322.jpg)
- 0
- 0
![Western blot - Anti-Nrf2 antibody [EP1808Y] (AB62352)](https://content.abcam.com/products/images/nrf2-antibody-ep1808y-ab62352--western-blot-img417623.jpg)
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 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.
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
Gas6 influences several cellular processes such as cell survival proliferation and migration. Gas6 exerts its effects in a complex with AXL receptor which is part of a larger network involving other TAM receptors. By activating AXL Gas6 modulates immune responses inflammation and tissue repair. The protein also plays a significant role in phagocytosis helping remove apoptotic cells thereby maintaining tissue homeostasis.
Pathways
The Gas6 protein is significantly engaged in the PI3K/AKT signaling pathway important for promoting cell survival and inhibiting apoptosis. Additionally Gas6 interacts with the NF-kB pathway which regulates inflammation and immune responses. Both pathways highlight the synergy between Gas6 and AXL emphasizing their contribution to cellular integrity and response to injury.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
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