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AB262109

Human MAP2K4 (MEK4/MKK4) knockout MCF7 cell line

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MAP2K4 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Sanger Sequencing - Human MAP2K4 (MEK4/MKK4) knockout MCF7 cell line (AB262109)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human MAP2K4 (MEK4/MKK4) knockout MCF7 cell line (AB262109)

Allele-1 : 1 bp deletion in exon1

Sanger Sequencing - Human MAP2K4 (MEK4/MKK4) knockout MCF7 cell line (AB262109)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human MAP2K4 (MEK4/MKK4) knockout MCF7 cell line (AB262109)

Allele-2 : 1 bp insertion in exon 1.

Key facts

Cell type

MCF7

Species or organism

Human

Tissue

Breast

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1

Disease

Adenocarcinoma

Reactivity data

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Product details

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

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?

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Properties and storage information

Gene name
MAP2K4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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
  • Slow to trypsinise.
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 5-7x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

MEM + 10% FBS + 0.01 mg/ml bovine insulin

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.

MEK4 also known as MKK4 is a protein kinase with a molecular weight of approximately 43 kDa. It functions as a dual-specificity kinase that phosphorylates and activates other kinases in cellular signaling pathways. MEK4 appears in various tissues including heart brain and skeletal muscle. It acts as an essential component in transmitting extracellular signals to downstream effectors and is involved in critical cellular processes like proliferation differentiation and apoptosis.
Biological function summary

MEK4 acts as an upstream kinase in the MAPK signaling cascade. Through its role in this pathway it is part of a larger signaling complex. MEK4 specifically activates JUN N-terminal kinase (JNK) by phosphorylation. JNK further regulates gene expression by influencing transcription factors impacting cellular responses. This makes MEK4 a pivotal element in facilitating messenger roles that allow cells to adapt to changing environments such as stress responses and immune reactions.

Pathways

MEK4 plays significant roles in the MAPK and stress-activated protein kinase (SAPK) pathways. In the MAPK pathway MEK4 phosphorylates JNK thereby influencing cellular responses to environmental changes. Within the SAPK pathway MEK4 interacts with other kinases such as MKK7 to modulate cellular responses further. The flow of signals through these pathways affects cell fate cell cycle regulation and programmed cell death demonstrating its comprehensive influence on cell physiology.

MEK4 shows involvement in cancer progression and neurodegenerative diseases. Abnormal activation of MEK4 can contribute to tumor growth and metastasis in cancers such as prostate and ovarian cancer by enhancing JNK signaling. In neurodegenerative disorders deregulation of MEK4-related pathways might lead to neuronal damage or apoptosis. Proteins like JNK and other MAP kinases become dysregulated in these conditions illustrating the interconnected role of MEK4 in disease mechanisms. Phospho flow cytometry can be utilized to measure alterations in MEK4 phosphorylation states aiding in elucidating its role in disease contexts.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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