Human MAP3K5 knockout A549 cell line
Be the first to review this product! Submit a review
|
(0 Publication)
- NGS
Lab
Next Generation Sequencing - Human MAP3K5 knockout A549 cell line (AB301060)
91 bp deletion in exon 1, CCDS5179.1
- WB
Lab
Western blot - Human MAP3K5 knockout A549 cell line (AB301060)
Western blot : Rabbit Monoclonal [EP553Y] to ASK1 ab45178 staining at 1/2000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 150-170 kDa in Wild-type A549 cell lysates with no signal observed at this size in MAP3K5 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-ASK1 antibody [EP553Y] (<a href='/en-us/products/primary-antibodies/ask1-antibody-ep553y-ab45178'>ab45178</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 at 20 µg
Lane 2:
Western blot - Human MAP3K5 knockout A549 cell line (ab301060) at 20 µg
Lane 3:
U-2 OS at 20 µg
Secondary
Lanes 1 - 3:
Goat anti-Rabbit 800CW at 1/20000 dilution
Lanes 1 - 3:
Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 155 kDa
Observed band size: 150-170 kDa,50 kDa
false
Complementary Products
![Flow Cytometry (Intracellular) - Anti-ASK1 antibody [EP553Y] (AB45178)](https://content.abcam.com/products/images/ask1-antibody-ep553y-ab45178--flow-cytometry-intracellular-img37728.jpg)
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab288558). 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.
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
In response to stressors ASK1 activates the c-Jun N-terminal kinase (JNK) and p38 MAPK signaling pathways which play roles in apoptosis and inflammation. ASK1 tends to form a complex with TRAF2 and TRAF6 key adapters in downstream signaling. This kinase acts as an important link bridging environmental stress signals to cellular responses. Through its function ASK1 influences cell fate decisions in response to various stress conditions.
Pathways
ASK1 is a central player in the MAPK signaling cascade. Within this cascade ASK1 regulates JNK and p38 MAPK pathways which are significant in stress adaptation and cellular responses. ASK1 interacts with proteins like MKK4 and MKK7 in the JNK pathway amplifying the stress signals. Through these interactions ASK1 ensures the propagation of signals important for adaptive responses or programmed cell death.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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