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Human KEAP1 Knockout Cell Line-A549
Human KEAP1 Knockout Cell Line-A549
Human KEAP1 Knockout Cell Line-A549

Human KEAP1 Knockout Cell Line-A549

Cat.No. :  CSC-RT2794

Host Cell:  A549 Target Gene:  KEAP1

Size:  1x10^6 cells/vial, 1mL Validation:  Sequencing

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Cell Line Information

Cell Culture Information

Safety and Packaging

Cat. No. CSC-RT2794
Description This cell is a stable cell line with a homozygous knockout of human KEAP1 using CRISPR/Cas9.
Target Gene KEAP1
Host Cell A549
Host Cell Species Homo sapiens (Human)
Size Form 1 vial (>10^6 cell/vial)
Shipping Dry ice package
Storage Liquid Nitrogen
Revival Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media.
Mycoplasma Negative
Format One frozen vial containing millions of cells
Storage Liquid nitrogen
Safety Considerations

The following safety precautions should be observed.

1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.

2. No eating, drinking or smoking while handling the stable line.

3. Wash hands after handling the stable line and before leaving the lab.

4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.

5. All waste should be considered hazardous.

6. Dispose of all liquid waste after each experiment and treat with bleach.

Ship Dry ice
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Background

Applications

Publications

Q & A

Customer Reviews

KEAP1 (Kelch-like ECH-associated protein 1) is a key gene involved in the cellular defense mechanism against oxidative stress. KEAP1 functions primarily as a negative regulator of NRF2. Under basal conditions, KEAP1 binds to NRF2, promoting its ubiquitination and subsequent proteasomal degradation. This interaction ensures that NRF2 levels remain low under normal physiological conditions. However, under the action of oxidative stress or electrophilic compounds, KEAP1 undergoes conformational changes that prevent its binding to NRF2, allowing NRF2 to accumulate and translocate into the nucleus. Once in the nucleus, NRF2 activates the expression of target genes involved in antioxidant defense, detoxification, and cell survival. Mutations and dysregulation of the KEAP1 gene are associated with various diseases, especially cancer. Loss-of-function mutations in KEAP1 can lead to uncontrolled activation of NRF2, resulting in excessive cell proliferation and survival. Such mutations are common in non-small cell lung cancer (NSCLC), liver cancer, and several other malignancies. Increased NRF2 activity provides cancer cells with a survival advantage, making them more resistant to chemotherapy and radiation therapy, posing a major challenge to cancer treatment. In addition, KEAP1 mutations have also been associated with neurodegenerative diseases, in which impaired oxidative stress responses may lead to neuronal damage.
Applications of Human KEAP1 Knockout Cell Line (A549) 1. Cancer Research: The KEAP1 knockout cell line (A549) is a valuable tool for studying the mechanisms of lung cancer. By knocking out the KEAP1 gene, researchers can investigate how the cellular pathways change, leading to cancer progression. This can lead to the identification of novel therapeutic targets specific to KEAP1 mutations or deletions. 2. Oxidative Stress Studies: KEAP1 is a crucial regulator of the antioxidant response. In KEAP1 knockout cells, the NRF2 pathway is constitutively activated. This makes this cell line an excellent model for studying oxidative stress and the cellular response to various oxidative agents, potentially leading to new insights into oxidative stress-related diseases. 3. Drug Screening: This cell line can be used in high-throughput drug screening to identify compounds that selectively target KEAP1-deficient cells. Such compounds can be promising candidates for the development of new cancer treatments, particularly for patients with KEAP1 mutations. 4. Pathway Dissection: With KEAP1 knocked out, it becomes possible to dissect the downstream effects and interactions within various cellular signaling pathways. This can shed light on how KEAP1 interacts with other proteins and pathways, contributing to a more comprehensive understanding of cellular homeostasis and response to stress. 5. Biomarker Discovery: By comparing the KEAP1 knockout cell line with the wild-type A549 cells, scientists can identify potential biomarkers that are specifically associated with the KEAP1-NRF2 pathway.
Customer Q&As
How is the knockout cell line validated?

A: The knockout cell product is validated by PCR amplification and Sanger Sequencing to confirm the mutation at the genomic level. Please find the detailed mutation info in the datasheet.

Is the product a single clonal cell or mixed cell pool?

A: Single clonal cell.

Can I confirm gene knockout by RT-qPCR?

A: No. This knockout cell product is generated using the CRISPR/Cas9 system to induce small insertions or deletions (indels) resulting in frameshift mutations. Although these frameshift mutations typically disrupt the coding gene, there is a possibility that the non-functional transcript may still be transcribed. Consequently, this could potentially yield misleading results when analyzed by RT-qPCR.

How can I store the cell product?

A: The cell line should be stored in liquid nitrogen for long-term preservation.

Is it possible to get multiple knockout clones for my GOI?

A: For most cases, we often keep at least 2 clones with different frameshift mutations. Please feel free to contact us to check if there are additional available clones.

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Customer Reviews
Clear, reproducible results

I have been using the Human KEAP1 Knockout Cell Line-A549 for over six months now and the results have been phenomenal. This cell line has provided clear, reproducible results in my oxidative stress experiments.

Germany

08/12/2021

Highly recommend

The KEAP1 Knockout Cell Line-A549 has dramatically accelerated our drug discovery process. The consistency and reliability of the knockout validation have given us high confidence in our data.

United States

01/11/2022

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