Human PEA15 knockout U-87 MG cell line
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human PEA15 knockout U-87 MG cell line (AB306860)
Western blot : PEA15 Monoclonal Antibody (4D2) abMA5-47430 staining at 1/1000 dilution, shown in black. A band was observed at 15 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in PEA15 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc BSA 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-Mouse HRP (H+L) at 1/20,000 dilution.
All lanes:
Western blot at 1/1000 dilution
Lane 1:
Wild-type U-87 MG at 20 µg
Lane 2:
Western blot - Human PEA15 knockout U-87 MG cell line (ab306860) at 20 µg
Lane 3:
MCF7 at 20 µg
Lane 4:
HEK-293 Nuclear at 20 µg
Secondary
All lanes:
Goat anti-Mouse HRP (H+L) at 1/20000 dilution
true
Exposure time: 720s
- NGS
Supplier Data
Next Generation Sequencing - Human PEA15 knockout U-87 MG cell line (AB306860)
65 bp deletion after Leu10 of the WT protein
Reactivity data
Product details
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
EMEM + 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
PEA15 plays pivotal roles in promoting cellular survival and regulating apoptosis within various cell types. It operates by modulating pathways like apoptosis and cell proliferation often forming complexes with other DED-containing proteins such as FADD and caspase-8. By interacting with these proteins PEA15 can maintain cellular stability and manage stress responses illustrating its significant contribution to cellular homeostasis.
Pathways
PEA15 is integral in mediating the extracellular signal-regulated kinase (ERK) and MAP kinase pathways. As PEA15 binds to ERK1/2 it retains these kinases in the cytoplasm impeding their translocation to the nucleus and therefore affecting the transcription of target genes. Furthermore its interactions with Ras-related proteins indicate its involvement in the regulation of cell growth and differentiation showing its broad influence on cellular signaling cascades.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
![Immunohistochemistry - Anti-PEA15 antibody [4D2] (AB277626)](https://content.abcam.com/products/images/pea15-antibody-4d2-ab277626--immunohistochemistry-img117601.jpg)
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