Human PRKCI (PKC iota) knockout HeLa cell line
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(0 Publication)
- WB
Lab
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] (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 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
false
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCI (PKC iota) knockout HeLa cell line (AB264670)
Homozygous : Insertion of the selection cassette in exon 5.
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Primary Antibodies
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (AB125066)](https://content.abcam.com/products/images/glutathione-peroxidase-4-antibody-epncir144-ab125066--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img436422.jpg)
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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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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