PKP3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon1.
PKP3_HUMAN, Plakophilin 3b, Plakophilin-3
PKP3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon1.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
Plakophilin 3 often referred to as PKP3 is a member of the Armadillo family of proteins. PKP3 is a desmosomal plaque protein with a known molecular mass of approximately 84 kDa. It localizes primarily to desmosomes and the nucleus where it contributes to cell-cell adhesion. PKP3 is commonly expressed in epithelial tissues where it helps maintain the structural integrity of epithelial sheets by binding to desmosomal cadherins.
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.
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.
Plakophilin 3 shows a connection to certain forms of cancer including prostate cancer and squamous cell carcinoma. PKP3 expression levels have an association with tumor progression and metastasis. In prostate cancer for instance altered expression of desmosomal components such as PKP3 can influence cancer cell adhesion and mobility. PKP3 interacts with proteins like E-cadherin in these pathological states which can affect adhesion properties and promote malignancy.
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False colour image of Western blot: Anti-Plakophilin 3 antibody [EPR5560] staining at 1/10000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-Plakophilin 3 antibody [EPR5560] 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 Human PKP3 (Plakophilin 3) knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Plakophilin 3 antibody [EPR5560] (Anti-Plakophilin 3 antibody [EPR5560] ab109441) 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 (ab258120)
Lane 2: Western blot - Human PKP3 (Plakophilin 3) knockout HeLa cell line (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
Homozygous: 5 bp deletion in exon1
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