PRKCI KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 5.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 5
Alternative names
Atypical protein kinase C-lambda/iota, DXS1179E, KPCI_HUMAN, MGC26534, PKC lambda, PKC lambda/iota, PKCI, PKCiota, PRKC iota, PRKC-lambda/iota, PRKCI, Protein kinase C iota, Protein kinase C iota type, aPKC-lambda/iota, nPKC-iota
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PRKCI KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 5.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 5
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- PRKCI
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
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
Protein Kinase C (PKC) iota also known by its gene symbol PRKCI is an atypical member of the PKC family. It has a molecular mass of approximately 74 kDa. PKC iota does not respond to calcium and is insensitive to diacylglycerol and phorbol esters which differentiates it from other classical PKC family members. The protein initiates cell survival and proliferation pathways and is primarily expressed in brain heart liver and skeletal muscle tissues indicating its extensive role across various biological functions.
- Biological function summary
PKC iota acts as an important player in regulating cell polarity and protecting cells from stress-induced apoptosis. It forms part of the Par6-Par3-PKC iota complex which is important for maintaining cell polarity and directional migration. This complex is essential for neural tube closure during embryonic development and in establishing cell polarity across diverse cell types. PKC iota also contributes to the regulation of microtubule organization reinforcing its vital role in maintaining cellular architecture.
- Pathways
PKC iota is important in the NF-kB signaling pathway and the PI3K/AKT pathway. In the NF-kB pathway PKC iota activates IkB kinase leading to NF-kB transcriptional activity which promotes cell survival. In the PI3K/AKT pathway PKC iota contributes to cell growth and survival working closely with AKT proteins. These pathways highlight its involvement in processes like proliferation survival and cellular response to external stimuli.
- Associated diseases and disorders
PKC iota has a significant connection to cancer and neurological disorders. Its overexpression has been observed in several cancers such as lung and prostate cancer where it is often associated with poor prognosis. PKC iota’s role in the maintenance of cell polarity and survival links it to malignant cell behavior. Additionally in neurological disorders like Alzheimer’s disease alterations in PKC iota activity have been related to synaptic dysfunctions. Connections to proteins such as AKT and NF-kB in these conditions highlight its potential as a therapeutic target.
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2 product images
Western blot - Human PRKCI (PKC iota) knockout HeLa cell line (ab264670)
False colour image of Western blot: Anti-PRKCI antibody staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, the antibody was shown to bind specifically to PRKCI. A band was observed at 72 kDa in wild-type HeLa cell lysates with no signal observed at this size in PRKCI knockout cell line ab264670 (knockout cell lysate Human PRKCI (PKC iota) knockout HeLa cell lysate ab258604). To generate this image, wild-type and PRKCI knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Performed under reducing conditions.
Sanger Sequencing - Human PRKCI (PKC iota) knockout HeLa cell line (ab264670)
Homozygous: Insertion of the selection cassette in exon 5.
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