Human MAPK7 (ERK5) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human MAPK7 (ERK5) knockout HeLa cell line (AB265508)
Lanes 1- 2 : Merged signal (red and green). Green - ab40809 observed at 115 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab40809 was shown to react with ERK5 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265508 (knockout cell lysate ab258042) was used. Wild-type HeLa and MAPK7 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40809 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ERK5 antibody [EP791Y] (<a href='/en-us/products/primary-antibodies/erk5-antibody-ep791y-ab40809'>ab40809</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MAPK7 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MAPK7 (ERK5) knockout HeLa cell line (ab265508)
Predicted band size: 88 kDa
Observed band size: 115 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MAPK7 (ERK5) knockout HeLa cell line (AB265508)
Homozygous : 1 bp insertion in exon 4.
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Primary Antibodies
AB138498
Anti-non-muscle Myosin IIA antibody [EPR8965]
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-non-muscle Myosin IIA antibody [EPR8965] (AB138498)](https://content.abcam.com/products/images/non-muscle-myosin-iia-antibody-epr8965-ab138498--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img71168.jpg)
- 5
- 1
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
Zygosity
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
ERK5 influences cellular functions through its role in promoting cell proliferation and differentiation. It serves as part of a signaling complex that includes the MAP kinase kinase 5 (MEK5). This interaction allows ERK5 to phosphorylate and subsequently activate target substrates such as transcription factors promoting cellular responses necessary for development and survival. ERK5 also plays a role in the response to oxidative stress which contributes to its impact on survival pathways and cellular stress responses.
Pathways
ERK5 acts within the MAPK signaling cascade and is linked to the ERK/MAPK pathway and the phosphoinositide 3-kinase (PI3K) pathway. Within these pathways ERK5 interacts with other MAP kinases and regulatory proteins such as Ras and Raf contributing to a wide range of cellular processes including growth migration and survival. Through these interactions ERK5 influences cellular dynamics and mediates responses to growth factors and stress stimuli.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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