Human PKP3 (Plakophilin 3) knockout HeLa cell line
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PKP3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 1.
View Alternative Names
PKP3_HUMAN, Plakophilin 3b, Plakophilin-3
- WB
Unknown
Western blot - Human PKP3 (Plakophilin 3) knockout HeLa cell line (AB265539)
False colour image of Western blot : Anti-Plakophilin 3 antibody [EPR5560] staining at 1/10000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab109441 was shown to bind specifically to Plakophilin 3. A band was observed at 70 and 80 kDa in wild-type HeLa cell lysates with no signal observed at this size in PKP3 CRISPR-Cas9 edited cell line ab265539 (CRISPR-Cas9 edited cell lysate ab258120). The band observed in the CRISPR-Cas9 edited lysate lane below 70 and 80 kDa is likely to represent a truncated form of Plakophilin 3. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and PKP3 CRISPR-Cas9 edited HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Plakophilin 3 antibody [EPR5560] (<a href='/en-us/products/primary-antibodies/plakophilin-3-antibody-epr5560-ab109441'>ab109441</a>) at 1/10000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PKP3 CRISPR-Cas9 edited HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PKP3 (Plakophilin 3) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-pkp3-plakophilin-3-knockout-hela-cell-lysate-ab258120'>ab258120</a>)
Lane 2:
Western blot - Human PKP3 (Plakophilin 3) knockout HeLa cell line (ab265539)
Lane 3:
A431 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Predicted band size: 87 kDa
Observed band size: 70 kDa,80 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PKP3 (Plakophilin 3) knockout HeLa cell line (AB265539)
Homozygous : 5 bp deletion in exon 1.
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
Plakophilin 3 plays a vital role in stabilizing junctional complexes by anchoring desmosomal components to the cytoskeleton. It usually forms a part of the desmosome a cellular structure composed of desmogleins desmocollins and plakoglobin. Through these interactions PKP3 contributes to the robust mechanical strength of tissue architecture and participates in signaling pathways that regulate cell proliferation and differentiation.
Pathways
Plakophilin 3 integrates into desmosome-related signaling pathways notably the intercellular adhesion pathways and the Wnt signaling pathway. PKP3 associates with proteins such as plakoglobin and β-catenin which are also involved in Wnt signaling influencing cell-cell contact and transcriptional regulation. These pathways are important for maintaining the balance between cell adhesion and cellular signaling ensuring appropriate responses to developmental and environmental cues.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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