anti-Perilipin 3 (mouse N-Terminus) guinea pig polyclonal, serum
- Guinea pig polyclonal
- Suitable for IHC and WB
- Reacts with mouse and 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|
|Immunogen||Synthetic peptide of murine TIP47 N-terminus (aa 1 – 14), MSSNGTDAPAEAQA-C|
|Formulation||Contains 0.09% sodium azide and 0.5% BSA|
|Stability||Guinea pig polyclonal (mNT)|
|UniprotID||Q9DBG5 (Mouse),M0RA08 (Rat)|
|Note||Centrifuge prior to opening|
|Tested applications||Tested dilutions|
|Immunohistochemistry (IHC) - frozen||1:100-1:200|
|Immunohistochemistry (IHC) - paraffin||1:100-1:200 (microwave treatment recommended)|
|Western Blot (WB)||1:2,000|
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 GP36 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)
Q & A's
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- 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.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
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.
- 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.
- 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.
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.