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anti-Perilipin 3 (mouse C-Terminus) guinea pig polyclonal, serum

Key Features
  •     Guinea pig polyclonal
  •     Suitable for IHC and WB
  •     Reacts with mouse and rat
Product description
Quantity 100 µl
Antibody Type Polyclonal
Host Guinea pig
Conjugate Unconjugated
Application IHC, WB
Purification Stabilized antiserum
Reactivity Mouse, Rat
No reactivity Human
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 Synthetic peptide of murine TIP47 C-terminus (aa 424 – 437), C-PFAPGITEKTPEGK
Formulation Contains 0.09% sodium azide and 0.5% BSA
UniprotID Q9DBG5 (Mouse),M0RA08 (Rat)
Note Centrifuge prior to opening
Applications
Tested applications Tested dilutions
Immunohistochemistry (IHC) - frozen 1:100-1:200
Western Blot (WB) 1:2,000
Background

TIP47 (tail-interacting protein of 47 kD, also named PLIN3) is involved in lipid droplet maturation. The protein has been localized on the surface of lipid droplets present in many cells and tissues, especially in milk fat globule membranes of human and bovine origin.
TIP47 shows about 40% sequence homology to adipophilin. The GP37 antiserum, however, is specific for TIP47/PLIN3 and does not cross-react with adipophilin and perilipin or other proteins of the PLIN/PAT family.
MW 47 262 (calculated from aa sequence data); apparent 52 000 (after SDS-PAGE); pI = 5.45

Reactivity on cultured cell lines: 3T3 and OP9 cells (mouse)

References/Publications (1)
Publication
Species
Application
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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.

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. 
MFGM (human or bovine) from milk
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.
The use of milk preparations (skim milk, dry milk) for blocking (and also for antibody dilution buffer) might be problematic. The antigens of Perilipin 3 antibodies were originally found and described in bovine milk. Some dry milk brands (or even different lots) might contain the epitopes and block it, thus abolishing the reaction with the antigen transferred onto the Western blot membrane and/or causing background staining on the transfer membrane itself. In this case Tween 20 or BSA or ovalbumin is recommended as e.g. blocking material.
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.

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