anti-Plakophilin 1 guinea pig polyclonal, serum

Cat. No. GP-PP1

Available, delivery time 1-7 days

€268.00*

Key Features
  • Guinea pig polyclonal
  • Suitable for IHC and WB
  • Reacts with bovine, human and mouse
Product description
Quantity 100 µl
Antibody Type Polyclonal
Host Guinea pig
Conjugate Unconjugated
Application IHC, WB
Purification Stabilized antiserum
Reactivity Bovine, Human, Mouse
Storage Short term at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles
Intended use Research use only
Immunogen Human recombinant plakophilin 1
Formulation Contains 0.09% sodium azide and 0.5% BSA
UniprotID Q28161 (Bovine), Q13835 (Human), P97350 (Mouse)
Synonym Plakophilin-1, Band 6 protein, B6P, PKP1
Note Centrifuge prior to opening
Applications
Tested applications Tested dilutions
Immunohistochemistry (IHC) - frozen 1:100
Western Blot (WB) 1:1,000
Background

PP1 antiserum shows distinct suprabasal staining of stratified epithelia, localizing plakophilin 1 in desmosomal plaques and in the nucleus. In Western blot analysis the antiserum reacts specifically with the 80 kDa plakophilin 1a and 1b polypeptides.

Reactivity on cultured cell lines: BPH-1 (derived from non-neoplastic prostatic tissue).

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FAQs
  • PVDF membranes show better results than nitrocellulose (higher capacity, allows for more stringent washing conditions in case of background problems).
  • Use freshly prepared blocking solution (e.g. 5% nonfat dry milk, 0.05% Tween 20), block for at least 1 h at room temperature.
  • Use the antibody in a higher dilution, but prolong incubation time and exposure time.
  • Always use a fresh aliquot of the antibody.
  • Do not repeatedly freeze the antibody (eventually centrifuge shortly after thawing to remove cryo-precipitates).
  • Include an additional washing step.
    You might also try more stringent wash conditions, e.g. add 0.5 M NaCl to the wash buffer.
  • Always use a fresh aliquot of secondary antibody.
  • In case you use ECL most the guinea pig antibody should be diluted further in order to get rid of the background.

Epidermis
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.

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.
  1. Methanol/ acetone fixation: Immerse slide in precooled (-20°C) methanol for 5 min, immerse in precooled (-20°C) acetone for 30-60 sec, let specimen air dry before antibody incubation.
  2. Methanol/ acetone fixation plus detergent permeabilization: After methanol/ acetone fixation and air-drying dip slide either in a solution containing 0.1-0.2% Triton X-100 in PBS or in 0.1% saponin in PBS for 1-5 min at room temperature (enhances accessibility of many cytoskeletal antigens).

  • Air-drying of the section.
  • Block with the serum of the species in which the secondary antibody was raised for 30 min.
  • Incubation with 1st antibody 1 h at RT in moist chamber.
  • Wash 3x with PBS.
  • Incubation with appropriate fluorescent secondary antibody, 30-60 min at RT.
  • Wash 3x with PBS.
  • Immerse shortly into ethanol.
  • Let air dry.
  • Cover with mounting medium.

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.

The concentration of specific antibody in our guinea pig serum is not determined.
In guinea pigs the antibody concentration in serum varies from 10 to 20 mg/ml; specific antibodies represent normally about 0.1-1% of total IgG. Total protein concentration varies from 40 to 65 mg/ml, with the main constituent (about 60%) being albumin.
  • 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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