anti-Plakophilin 2 mouse monoclonal, PP2/62 + PP2/86 + PP2/150, supernatant

Supernatant

5 ml

Cat. No. 651101

Prices excl. VAT

Available, delivery time: 1-7 days

€228.00*

Datasheet

Key Features

Hybridoma cell culture supernatant
Mouse monoclonal
Suitable for IHC, IP and WB
Reacts with bovine, human and rat
Isotype IgG1

Product description

Quantity	5 ml
Antibody Type	Monoclonal
Host	Mouse
Isotype	IgG1
Conjugate	Unconjugated
Application	IHC, WB
Purification	Hybridoma cell culture supernatant
Reactivity	Human, Rat
Storage	Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles
Intended use	Research use only
Clone	PP2/62 + PP2/86 + PP2/150
Immunogen	Synthetic carboxyterminal peptide (24 aa) of human plakophilin 2
Formulation	Contains 0.09% sodium azide
UniprotID	A0A3Q1M2G3 (Bovine), Q99959 (Human), Q562C0 (Rat)
Synonym	Plakophilin-2, PKP2

Applications

Tested applications	Tested dilutions
Immunohistochemistry (IHC) - frozen	1:2-1:50 (incubate for 5 min in PBS plus 0.2% Triton-X100 prior to antibody application; longer incubation could result in a loss of nuclear staining; fixation with 2% formaldehyde is recommended; exclusive nuckear staining without detergent)
Immunohistochemistry (IHC) - paraffin	1:2-1:50 (microwave treatment recommended)
Western Blot (WB)	1:50-1:100

Background

The mabs react with plakophilin 2, a member of the arm-repeat family of proteins which can occur in at least 2 splice forms (2a and 2b). Plakophilin 2 assembles with other proteins to form the desmosomal plaque in simple and glandular epithelia, the basal layer of certain stratified epithelia, all layers of some stratified epithelia, and in nonepithelial desmosome-possessing tissues such as myocardium, Purkinje fibers, and dendritic reticulum of lymph node follicles. In all these cells, including a variety of cell types devoid of desmosomes, plakophilin 2 is highly enriched in the karyoplasm. Polypeptide reacting: Plakophilin 2, Mr 100 kDa (SDS-PAGE); MW calculated from aa sequence: 92,750 (pI 9.33) and 97,410 (pI 9.38).

Tumors specifically detected: positive staining was obtained with several carcinomas derived from simple-type epithelia (e.g. hepatomas, colon carcinomas) and some adenocarcinomas derived from stratified epithelia (oesophagus carcinoma). The antibody was negative with all leiomyosarcomas tested.

References/Publications (27)

Publication

Species

Application

Publication Dietrich, N. et al. Adhesion of pancreatic tumor cell clusters by desmosomal molecules enhances early liver metastases formation. Sci Rep 14, 18189 (2024).

Species human, mouse

Application WB

Publication De Bortoli, M. et al. Modeling incomplete penetrance in arrhythmogenic cardiomyopathy by human induced pluripotent stem cell derived cardiomyocytes., Comput Struct Biotechnol J 21,1759-1773, (2023).

Species human

Application ICC-IF, WB

Publication Wanuske, M. T. et al. Clustering of desmosomal cadherins by desmoplakin is essential for cell-cell adhesion., Acta Physiol (Oxf) 231, e13609, (2021).

Species human

Application WB

Publication Janz A. et al. CRISPR/Cas9-edited PKP2 knock-out (JMUi001-A-2) and DSG2 knock-out (JMUi001-A-3) iPSC lines as an isogenic human model system for arrhythmogenic cardiomyopathy (ACM)., Stem Cell Res, 53, 102256, (2021).

Species human

Application IHC/IF, WB, FACS

Publication Hamada, Y. et al. G790del mutation in DSC2 alone is insufficient to develop the pathogenesis of ARVC in a mouse model. Biochem Biophys Rep. 21, 100711(2020).

Species mouse

Application IHC/IF, WB

Publication Merkel, C. et al. Vinculin anchors contractile actin to the cardiomyocyte adherens junction. Mol.Biol.Cell. 30, 2639-2650 (2019)

Species mouse

Application IHC (paraffin)

Publication Domke, L and Franke, W. The cell-cell junctions of mammalian testes.., Cell Tissue Res, 375, 451-482, (2019)

Species bovine

Application ICC-IF

Publication Li, Y. et al. The N-cadherin interactome in primary cardiomyocytes as defined using quantitative proximity proteomics. J.Cell.Sci. 132, (2019)

Species mouse

Application ICC-IF

Publication Gergs, U. et al. Alterations of protein expression of phospholamban, ZASP and plakoglobin in human atria in subgroups of seniors. Sci.Rep. 9, 5610 (2019)

Species human

Application WB

Publication Kam, C. et al. Desmoplakin maintains gap junctions by inhibiting Ras/MAPK and lysosomal degradation of connexin-43. J.Cell.Biol. 217, 3219-3235 (2018)

Species mouse

Application WB

Publication Niell, N. et al. The human PKP2/plakophilin-2 gene is induced by Wnt/β-catenin in normal and colon cancer-associated fibroblasts. Int.J.Cancer. 142, 792-804 (2018).

Species human

Application WB

Publication Dubash, A. D. et al. Plakophilin-2 loss promotes TGF-β1/p38 MAPK-dependent fibrotic gene expression in cardiomyocytes. J. Cell Biol. 212, 425–38 (2016).

Species rat

Application WB,ICC-IF

Publication Kant, S., Holthöfer, B., Magin, T. M., Krusche, C. A. & Leube, R. E. Desmoglein 2-Dependent Arrhythmogenic Cardiomyopathy Is Caused by a Loss of Adhesive Function. Circ. Cardiovasc. Genet. 8, 553–63 (2015).

Species mouse

Application IHC (paraffin)

Publication Rappe, U., Schlechter, T., Aschoff, M., Hotz-Wagenblatt, A. & Hofmann, I. Nuclear ARVCF Protein Binds Splicing Factors and Contributes to the Regulation of Alternative Splicing. J. Biol. Chem. 289, 12421–12434 (2014).

Species human

Application WB

Publication Patel, D. M., Dubash, A. D., Kreitzer, G. & Green, K. J. Disease mutations in desmoplakin inhibit Cx43 membrane targeting mediated by desmoplakin-EB1 interactions. J. Cell Biol. 206, 779–97 (2014).

Species human

Application ICC-IF

Publication Fischer-Kešo, R. et al. Plakophilins 1 and 3 bind to FXR1 and thereby influence the mRNA stability of desmosomal proteins. Mol. Cell. Biol. 34, 4244–56 (2014).

Species human

Application WB,IP

Publication Pieperhoff, S. et al. The plaque protein myozap identified as a novel major component of adhering junctions in endothelia of the blood and the lymph vascular systems. J. Cell. Mol. Med 16, 1709–1719 (2012).

Species mouse

Application WB

Publication Rickelt, S. Plakophilin-2: a cell-cell adhesion plaque molecule of selective and fundamental importance in cardiac functions and tumor cell growth. Cell Tissue Res. 348, 281–94 (2012).

Species human,mouse

Application WB,IHC (paraffin)

Publication Fabritz, L. et al. Load-reducing therapy prevents development of arrhythmogenic right ventricular cardiomyopathy in plakoglobin-deficient mice. J. Am. Coll. Cardiol. 57, 740–750 (2011).

Species mouse

Application IHC

Publication Rickelt, S., Winter-Simanowski, S., Noffz, E., Kuhn, C. & Franke, W. W. Upregulation of plakophilin-2 and its acquisition to adherens junctions identifies a novel molecular ensemble of cell-cell-attachment characteristic for transformed mesenchymal cells

Species human

Application WB,ICC-IF

Publication Goossens, S. et al. A unique and specific interaction between T-catenin and plakophilin-2 in the area composita, the mixed-type junctional structure of cardiac intercalated discs. J. Cell Sci. 120, 2126–2136 (2007).

Species mouse

Application WB,IHC (paraffin),IEM

Publication Pieperhoff, S. & Franke, W. W. The area composita of adhering junctions connecting heart muscle cells of vertebrates - IV: coalescence and amalgamation of desmosomal and adhaerens junction components - late processes in mammalian heart development. Eur.

Species mouse

Application IHC (frozen)

Publication Koeser, J., Troyanovsky, S. M., Grund, C. & Franke, W. W. De novo formation of desmosomes in cultured cells upon transfection of genes encoding specific desmosomal components. Exp. Cell Res. 285, 114–30 (2003).

Species human

Application WB,ICC-IF

Publication Akat, K. et al. Molecular characterization of desmosomes in meningiomas and arachnoidal tissue. Acta Neuropathol. 106, 337–347 (2003).

Species human

Application WB

Publication South, A. P. et al. Lack of plakophilin 1 increases keratinocyte migration and reduces desmosome stability. J. Cell Sci. 116, 3303–3314 (2003).

Species human

Application WB

Publication Mertens, C., Kuhn, C., Moll, R., Schwetlick, I. & Franke, W. W. Desmosomal plakophilin 2 as a differentiation marker in normal and malignant tissues. Differentiation 64, 277–290 (1999).

Species human

Application IHC (frozen),ICC-IF

Publication Mertens, C., Kulm, C. & Franke, W. W. Plakophilins 2a and 2b: Constitutive Proteins of Dual Location in the Karyoplasm and the Desmosomal Plaque. J. Cell Biol. 135, 1009–1025 (1996).

Species human

Application IHC (frozen),ICC-IF

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Datasheet

MSDS

FAQs

The concentration of unpurified supernatant, ascites, unpurified guinea pig serum and unpurified rabbit serum is not determined.
The concentration of purified antibodies is mentioned on the datasheet.
For prediluted antibodies the concentration may vary from lot to lot. The concentration of these antibodies is not mentioned on the datasheet and can be requested at support@progen.com.

Most of our purified mouse antibodies contain 0.5% BSA as stabilizer. If BSA was added to the antibody solution, it is stated in the datasheet.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.

We recommend heat induced antigen retrieval with 100 mM Tris-HCl (pH 9.5), 5% urea, 10 min at 120°C.

PROGEN offers different formats for mouse monoclonal antibodies. Different antibodies may not be offered in each format, the following are all possible formats:

Supernatant and supernatant concentrate: This format contains hybridoma cell culture supernatant. The antibody is not purified and the antibody concentration is not determined. The antibody concentration may vary from lot to lot. Therefore we recommend to titrate the optimal concentration for the application used for each new lot.
Lyophilized, purified: This format contains purified antibody in lyophilized form. The reconstitution of this antibody is described in the datasheet. The buffer composition after reconstitution is also mentioned on the datasheet.
Liquid, purified: This format contains purified antibody in liquid format. The concentration is mentioned on the datasheet.
Prediluted, purified: This format contains purified antibody in liquid format. Most antibodies in this format are diluted to be ready-to-use for IHC with standard tissue. But some antibodies of this format need further dilution for IHC. This is mentioned on the datasheet.

Lyophilized antibodies can be stored at 2-8°C until expiration.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.

The expiration date of our antibodies is indicated on the product label.

We offer bulk amounts starting from 1 mg for many of our mouse monoclonal or recombinant antibodies. Please contact us for further information or use the “bulk button” on the product page.

Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.

The optimal antibody dilution for your specific protocol and application needs to be titrated in your lab with your equipment and sample. The optimal dilution may vary between protocols and samples. A good dilution for starting the titration is the dilution mentioned in the datasheet. If the sample needs a specific treatment (eg. Antigen retrieval for IHC on FFPE sections) this should also be mentioned on the antibody datasheet.

PROGEN antibodies are shipped at ambient temperature. The antibodies are stable at ambient temperature for the shipment period. Please store the antibodies as indicated in the datasheet upon arrival.

Homogenization of tissue in PBS (supplemented with protease inhibitors).
Centrifugation at 10E4 x g.
Optional: treat pellet with DNase.
Resuspend pellet in detergent buffer (e.g. 1% Triton X-100), stir for 1 h at 2-8°C.
Centrifugation at 10E4 x g.
Resuspend pellet in high salt buffer (with 1.5 M KCl), stir for 1 h at 2-8°C.
Centrifugation at 10E4 x g.
Resuspend pellet in low salt buffer (e.g. 50 mM NaCl), stir for 30 min at 2-8°C.
Centrifugation at 10E4 x g: pellet contains raw cytoskeletal fraction.

Transfer from a low concentration acrylamide gel (e.g. 8%).
Reduce the methanol concentration of the transfer buffer (limit is 50% of the routine concentration 20% methanol).
Use the wet transfer method and transfer at least 4 h (or better overnight) at 100-200 mA.
In order to prevent the accumulation of gas bubbles, stir the transfer buffer continuously during the transfer (e.g. with a magnetic stirrer).
If possible use a cytoskeletal preparation of your material to be analyzed (in the case of cultured cells additionally DNase treatment) rather than whole cell lysate.
As a general rule we would like to recommend loading that much protein into one gel lane that the intermediate filament polypeptides (in the 40-60 kD range) can be readily detected in a control gel stained with Coomassie blue.

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