PMAIP1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1
Alternative names
APR_HUMAN, ATL-derived, Adult T cell leukemia derived PMA responsive, Immediate-early-response protein APR, NOXA, PMA-induced protein 1, PMA-responsive gene, Phorbol-12-myristate-13-acetate-induced protein 1, Pmaip1, Protein Noxa
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PMAIP1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 1 and 2 bp deletion in exon 1
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- PMAIP1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Noxa also known as PMAIP1 is a pro-apoptotic member of the Bcl-2 protein family. It has a molecular weight of approximately 20 kDa. Noxa commonly expresses in a number of tissues including lymphoid and myeloid lineages. It functions by interacting with other Bcl-2 family proteins like Mcl-1 promoting apoptosis through binding and neutralization of anti-apoptotic proteins. This interaction facilitates the release of cytochrome c from the mitochondria triggering caspase activation and subsequent apoptosis.
- Biological function summary
Noxa plays a role in the regulation of programmed cell death including being part of multiprotein complexes within the mitochondria. Noxa's expression is upregulated in response to various cellular stress signals. These stresses include DNA damage and hypoxia where it acts to mediate apoptosis preventing the survival of cells with potential mutations. Its role in modulating apoptosis makes it key in maintaining cellular homeostasis.
- Pathways
Noxa is involved in significant apoptotic and cellular stress response pathways. It forms part of the intrinsic apoptosis pathway which is tightly regulated by Bcl-2 family members like BAK and BAX. Noxa's ability to neutralize Mcl-1 allows the activation of these proteins committing the cell to apoptosis. It also participates in the p53 signaling pathway which is important for the cellular response to DNA damage wherein p53 directly transactivates Noxa.
- Associated diseases and disorders
Noxa is important in cancer and autoimmune syndromes. In cancer its expression often correlates with the sensitivity of cancer cells to chemotherapy as its role in apoptosis can enhance the elimination of cancerous cells. Mutations in p53 that impair its function can lead to decreased Noxa expression and apoptosis contributing to cancer progression. In autoimmune disorders aberrant regulation of apoptosis sometimes involving Noxa and its interaction with Bcl-2 proteins can result in premature cell death or survival of autoreactive cells exacerbating the disease.
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3 product images
Cell Culture - Human PMAIP1 (Noxa) knockout A549 cell line (ab267135)
Representative images of PMAIP1 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Sanger Sequencing - Human PMAIP1 (Noxa) knockout A549 cell line (ab267135)
Allele-2: 1 bp insertion in exon 1.
Sanger Sequencing - Human PMAIP1 (Noxa) knockout A549 cell line (ab267135)
Allele-1: 2 bp deletion in exon1
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