- 32-5700 - Invitrogen Antibodies PKP1 antibody

Submitted references Analysis of buccal mucosa as a prognostic tool in children with arrhythmogenic cardiomyopathy.

Bueno-Beti C, Field E, Tsatsopoulou A, Perry G, Sheppard MN, Behr ER, Saffitz JE, Kaski JP, Asimaki A
Progress in pediatric cardiology 2022 Mar;64:None

Homozygous splice site mutations in PKP1 result in loss of epidermal plakophilin 1 expression and underlie ectodermal dysplasia/skin fragility syndrome in two consanguineous families.

Sprecher E, Molho-Pessach V, Ingber A, Sagi E, Indelman M, Bergman R
The Journal of investigative dermatology 2004 Mar;122(3):647-51

Defining desmosomal plakophilin-3 interactions.

Bonné S, Gilbert B, Hatzfeld M, Chen X, Green KJ, van Roy F
The Journal of cell biology 2003 Apr 28;161(2):403-16

Desmoglein 4 in hair follicle differentiation and epidermal adhesion: evidence from inherited hypotrichosis and acquired pemphigus vulgaris.

Kljuic A, Bazzi H, Sundberg JP, Martinez-Mir A, O'Shaughnessy R, Mahoney MG, Levy M, Montagutelli X, Ahmad W, Aita VM, Gordon D, Uitto J, Whiting D, Ott J, Fischer S, Gilliam TC, Jahoda CA, Morris RJ, Panteleyev AA, Nguyen VT, Christiano AM
Cell 2003 Apr 18;113(2):249-60

Desmoglein 4 in hair follicle differentiation and epidermal adhesion: evidence from inherited hypotrichosis and acquired pemphigus vulgaris.

Kljuic A, Bazzi H, Sundberg JP, Martinez-Mir A, O'Shaughnessy R, Mahoney MG, Levy M, Montagutelli X, Ahmad W, Aita VM, Gordon D, Uitto J, Whiting D, Ott J, Fischer S, Gilliam TC, Jahoda CA, Morris RJ, Panteleyev AA, Nguyen VT, Christiano AM
Cell 2003 Apr 18;113(2):249-60

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PKP1 was achieved by transfecting HeLa cells with PKP1 specific siRNAs (Silencer® select Product #s10581, s10582). Western blot analysis (Fig a) was performed using whole cell extracts from the PKP1 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and un-transfected cells (lane 1). The blots were probed with Anti-PKP1 Mouse monoclonal Antibody (Product # 37-6200, 2 µg/mL) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to PKP1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of A-431 (Lane 1) and PC-3 (Lane 2). The blot was probed with Anti-Plakophilin-1 Mouse Monoclonal Antibody (Product # 32-5700, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 80 kDa band corresponding to Plakophilin-1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Plakophilin (PKP1) was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PKP1 (10B2) Mouse Monoclonal Antibody (Product # 32-5700) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cell junctional localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. Immunodetection of PKPs using different mouse mAbs. PKP3, detected with mAb 23E3/4 (a and b), 12B11F8 (c), or PKP2 (d) show a similar expression pattern along cell-cell contacts in methanol-fixed HaCaT (a) and HCT8/E8 (b-d) cells. Larger magnifications display the punctate localization of these proteins along cell-cell contacts (a'-d'). Bar, 10 mum. (e) Western blot detection of PKP3 (lane 1, mAb 12B11F8; lane 2, 23E3/4), PKP2 (lane 3), and PKP1 (lane 4) in HaCaT protein lysates. Equal amounts of protein were loaded in each lane.

32-5700

Antibody data