anti-Plakophilin 2 mouse monoclonal, Pkp2-518, supernatant

Cat. No. 651167

Available, delivery time: 1-7 days

€245.00*

Key Features

Mouse monoclonal
Suitable for IEM, 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	IEM, IHC, IP, WB
Purification	Hybridoma cell culture supernatant
Reactivity	Human, Mouse, Rat
No reactivity	Amphibia, Fish
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	Pkp2-518
Immunogen	Synthetic carboxyterminal peptide (aa 527 - 872) of human plakophilin 2
Formulation	Contains 0.09% sodium azide
UniprotID	A0A3Q1M2G3 (Bovine), Q99959 (Human), Q9CQ73 (Mouse), Q562C0 (Rat)
Synonym	Plakophilin-2, PKP2

Applications

Tested applications	Tested dilutions
Immunohistochemistry (IHC) - frozen	Ready-to-use
Immunohistochemistry (IHC) - paraffin	Ready-to-use (microwave treatment recommended)
Cell-based Assay	Assay dependent
Immunoprecipitation (IP)	Assay dependent
Western Blot (WB)	1:5-1:10

Background

The mab recognizes plakophilin 2 in both locations: in the desmosomal plaque and in the karyoplasm. Plakophilin 2 is 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, in Merkel cells, and desmosome-possessing nonepithelial cells 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 nuclei (karyoplasm).
Polypeptide reacting: Plakophilin 2, M_r ~100 kDa (SDS-PAGE); MW calculated from aa sequence: 92,750 (pI 9.33) and 97,410 (pI 9.38)
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).
Recommendation for immunohistochemistry on frozen sections: For better accessibility of the desmosome-associated antigen frozen sections might be pre-incubated for 5 min in PBS plus 0.2% Triton X-100 prior to antibody application. Longer incubation could result in a loss of Pkp2-nuclear staining. To prevent wash-out of soluble forms of the antigen, fixation with 2% formaldehyde is recommended.

References/Publications (6)

Publication

Species

Application

Publication Shoykhet, M., Waschke, J. & Yeruva, S. Cardiomyocyte cohesion is increased after ADAM17 inhibition. Front Cell Dev Biol 11, (2023).

Species mouse

Application WB

Publication Shoykhet, M. et al. EGFR inhibition leads to enhanced desmosome assembly and cardiomyocyte cohesion via ROCK activation. JCI Insight 8, (2023).

Species mouse

Application WB

Publication Yeruva, S. et al. Cholinergic signaling impairs cardiomyocyte cohesion. Acta Physiologica 236, (2022).

Species mouse

Publication Ragni, C. V. et al. Amotl1 mediates sequestration of the Hippo effector Yap1 downstream of Fat4 to restrict heart growth. Nat. Commun. 8, 1–11 (2017)

Species mouse

Application ICC-IF

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),ICC-IF

Publication Rickelt, S. et al. A novel kind of tumor type-characteristic junction: plakophilin-2 as a major protein of adherens junctions in cardiac myxomata. Mod. Pathol. 23, 1429–1437 (2010).

Species human

Application WB,IHC (paraffin),ICC-IF

Downloads

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.

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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