GP40 | Guinea Pig Polyclonal Anti-Perilipin 2 (N-terminus aa 1-29)

anti-Perilipin 2 (N-terminus aa 1-29) guinea pig polyclonal, serum

Key Features

Stabilized antiserum
Guinea pig polyclonal
Suitable for ICC/IF, IHC and WB
Reacts with human

Product description

Quantity	100 µl
Antibody Type	Polyclonal
Host	Guinea pig
Conjugate	Unconjugated
Application	ICC/IF, IHC, WB
Purification	Stabilized antiserum
Reactivity	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	Synthetic peptide (N-terminal aa 1-29 of human and murine adipophilin)
Formulation	Contains 0.09% sodium azide
UniprotID	Q99541 (Human), P43883 (Mouse)
Synonym	Perilipin-2, Adipophilin, Adipose differentiation-related protein, ADRP, PLIN2, ADFP
Note	Centrifuge prior to opening

Applications

Tested applications	Tested dilutions
Immunocytochemistry (ICC)/ Immunofluorescence (IF)	1:100-1:200
Immunohistochemistry (IHC) - paraffin	1:100-1:500 (microwave treatment recommended)
Western Blot (WB)	1:500-1:1,000

Background

Perilipin 2/Adipophilin/ADRP/PLIN2 (a member of the PLIN/PAT family) is a ubiquitous component of lipid droplets. It has been found in milk fat globule membranes and on the surface of lipid droplets in various cultured cell lines; inducible by etomoxir.
Enhanced expression of Perilipin 2/Adipophilin/ADRP/PLIN2 is a useful marker for pathologies characterized by increased lipid droplet accumulation. Such diseases include atheroma, steatosis, obesity and certain cases of liposarcoma. It also seems to be a potent marker for atherosclerosis.
ADRP can also be used to study the virus entry via lipid droplets.
Polypeptide reacting: specific for Perilipin 2/Adipophilin/ADRP/PLIN2, MW 48,100 (calculated from aa sequence data); apparent Mr 52,000 (after SDS-PAGE); pI 6.72.
Immunolocalization: Adipophilin/PLIN2 is positively detected in the glandular cells of lactating mammary gland (ductal cells are negative), zona fasciculata of the adrenal gland, Sertoli cells of the testis, and in fat-accumulating hepatocytes of alcoholic cirrhotic fatty liver; adipocytes are negative.
Also positively stained are lipid-storing CD 68-positive macrophages.

References/Publications (44)

Publication

Species

Application

Publication Kaushik S. et al. Chaperone-mediated autophagy regulates adipocyte differentiation., Sci Adv, 8, eabq2733, (2022).

Species mouse

Application ICC/IF, WB

Publication Scantlebery, A. et al. The dysregulation of metabolic pathways and induction of the pentose phosphate pathway in renal ischaemia-reperfusion injury. J Pathol. 253, 404-414(2021).

Species mouse

Application IHC (paraffin)

Publication Gouna, G. et al. TREM2-dependent lipid droplet biogenesis in phagocytes is required for remyelination. J. Exp. Med. 218, (2021).

Species Mouse

Application IHC-Fr-IF, WB

Publication Kuramoto, K. et al.. The autophagy protein Becn1 improves insulin sensitivity by promoting adiponectin secretion via exocyst binding.. Cell Rep. 35, 109184(2021).

Species mouse

Application WB

Publication Maya-Monteiro, C. M. et al. Lipid Droplets Accumulate in the Hypothalamus of Mice and Humans with and without Metabolic Diseases. Neuroendocrinology 111, 263–272 (2021).

Species Human

Application IHC-IF

Publication Zhao, L. et al. TDP-43 facilitates milk lipid secretion by post-transcriptional regulation of Btn1a1 and Xdh. Nat.Commun. 11, 341 (2020)

Species mouse

Application IHC-IF (paraffin)

Publication Wagner, C. et al. Lysosomal acid lipase is the major acid retinyl ester hydrolase in cultured human hepatic stellate cells but not essential for retinyl ester degradation. Biochim.Biophys.Acta.Mol.Cell.Biol.Lipids. 1865, 158730 (2020)

Species human

Application WB

Publication Tadepalle, N. et al. Microtubule-dependent and independent roles of spastin in lipid droplet dispersion and biogenesis. Life.Sci.Alliance. 3, (2020)

Species mouse

Application WB,ICC-IF

Publication Lundåsen, T. et al. The PPAR pan-agonist tetradecylthioacetic acid promotes redistribution of plasma cholesterol towards large HDL. PLoS.One. 15, e0229322 (2020)

Species mouse

Application IHC (paraffin)

Publication Losier, T. et al. AMPK Promotes Xenophagy through Priming of Autophagic Kinases upon Detection of Bacterial Outer Membrane Vesicles. Cell.Rep. 26, 2150-2165.e5 (2019)

Species human,mouse

Application WB

Publication Sané, A. et al. SAR1B GTPase is necessary to protect intestinal cells from disorders of lipid homeostasis, oxidative stress, and inflammation. J.Lipid.Res. 60, 1755-1764 (2019)

Species human

Application WB

Publication Daemen, S. et al. Distinct lipid droplet characteristics and distribution unmask the apparent contradiction of the athlete's paradox. Mol.Metab. 17, 71-81 (2018).

Species human

Application WB

Publication Sundberg, J. et al. Sebaceous gland abnormalities in fatty acyl CoA reductase 2 (Far2) null mice result in primary cicatricial alopecia. PLoS.One. 13, e0205775 (2018).

Species mouse

Application IHC (frozen)

Publication Whytock, K. et al. Hormone-sensitive lipase preferentially redistributes to lipid droplets associated with perilipin-5 in human skeletal muscle during moderate-intensity exercise. J.Physiol. 596, 2077-2090 (2018)

Species human

Application IHC-IF (frozen)

Publication Krahmer, N. et al. Organellar Proteomics and Phospho-Proteomics Reveal Subcellular Reorganization in Diet-Induced Hepatic Steatosis. Dev.Cell. 47, 205-221.e7 (2018)

Species mouse

Application IHC-IF (frozen)

Publication Homma, T. et al. Double Knockout of Peroxiredoxin 4 (Prdx4) and Superoxide Dismutase 1 (Sod1) in Mice Results in Severe Liver Failure. Oxid.Med.Cell.Longev. 2018, 2812904 (2018).

Species mouse

Application ICC-IF

Publication Gemmink, A. et al. Lipid droplet dynamics and insulin sensitivity upon a 5-day high-fat diet in Caucasians and South Asians. Sci. Rep. 7, 1–9 (2017).

Species human

Application WB

Publication Yang, Y. et al. Angiotensin II induces cholesterol accumulation and injury in podocytes. Sci.Rep. 7, 10672 (2017).

Species human

Application ICC-IF

Publication Maier, A. et al. Hypoxia-inducible protein 2 Hig2/Hilpda mediates neutral lipid accumulation in macrophages and contributes to atherosclerosis in apolipoprotein E-deficient mice. FASEB.J. 31, 4971-4984 (2017)

Species mouse

Application WB

Publication Minami, S. et al. Lipophagy maintains energy homeostasis in the kidney proximal tubule during prolonged starvation. Autophagy. 13, 1629-1647 (2017)

Species mouse

Application WB

Publication Heier, C. et al. The phospholipase PNPLA7 functions as a lysophosphatidylcholine hydrolase and interacts with lipid droplets through its catalytic domain. J.Biol.Chem. 292, 19087-19098 (2017).

Species grivet monkey

Application WB

Publication Furukawa, S., Nagaike, M. & Ozaki, K. Databases for technical aspects of immunohistochemistry. J. Toxicol. Pathol. 30, 79-107 (2017).

Species mouse

Application IHC (paraffin)

Publication Kaushik, S. & Cuervo, A. M. AMPK-dependent phosphorylation of lipid droplet protein PLIN2 triggers its degradation by CMA. Autophagy 12, 432-438 (2016).

Species mouse

Application WB,ICC-IF

Publication Gallardo-Montejano, V. I. et al. Nuclear Perilipin 5 integrates lipid droplet lipolysis with PGC-1a/SIRT1-dependent transcriptional regulation of mitochondrial function. Nat. Commun. 7, (2016).

Species mouse

Application PLA,WB,ICC-IF

Publication Kozusko, K. et al. Clinical and molecular characterization of a novel PLIN1 frameshift mutation identified in patients with familial partial lipodystrophy Europe PMC Funders Group. Diabetes 64, 299-310 (2015).

Species human,mouse

Application WB

Publication Kaushik, S. & Cuervo, A. M. Degradation of lipid droplet-associated proteins by chaperone- mediated autophagy facilitates lipolysis. Nat. Cell Biol. 17, 759-770 (2015).

Species mouse

Application WB,ICC-IF

Publication Kuramoto, K. et al. Deficiency of a Lipid Droplet Protein, Perilipin 5, Suppresses Myocardial Lipid Accumulation, Thereby Preventing Type 1 Diabetes- Induced Heart Malfunction. Mol. Cell. Biol. -34, 2721-2731 (2014).

Species mouse

Application WB,IHC (frozen)

Publication Camus, G. et al. The hepatitis C virus core protein inhibits adipose triglyceride lipase (ATGL)-mediated lipid mobilization and enhances the ATGL interaction with comparative gene identification 58 (CGI-58) and lipid droplets. J. Biol. Chem. 289, 35770-8

Species human

Application WB

Publication Fernandez-Rojo, M. A. et al. Caveolin-1 Is Necessary for Hepatic Oxidative Lipid Metabolism: Evidence for Crosstalk between Caveolin-1 and Bile Acid Signaling. Cell Rep. 4, 238-247 (2013).

Species mouse

Application WB

Publication Straub, B. K. et al. Adipophilin/perilipin-2 as a lipid droplet-specific marker for metabolically active cells and diseases associated with metabolic dysregulation. Histopathology 62, 617-631 (2013).

Species human

Application IHC (frozen)

Publication Dahlhoff, M. et al. PLIN2, the major perilipin regulated during sebocyte differentiation, controls sebaceous lipid accumulation in vitro and sebaceous gland size in vivo NIH Public Access. Biochim. Biophys. Acta 1830, 4642-4649 (2013).

Species human,mouse

Application WB,IHC (paraffin)

Publication Pauloin, A. & Chanat, E. Prolactin and epidermal growth factor stimulate adipophilin synthesis in HC11 mouse mammary epithelial cells via the PI3-kinase/Akt/mTOR pathway. Biochim. Biophys. Acta 1823, 987-996 (2012).

Species mouse

Application WB

Publication Bosma, M. et al. The lipid droplet coat protein perilipin 5 also localizes to muscle mitochondria. Histochem Cell Biol 137, 205-216 (2012).

Species rat

Application WB

Publication Lenz, L.-S. et al. Adipocyte-specific Inactivation of Acyl-CoA Synthetase Fatty Acid Transport Protein 4 (Fatp4) in Mice Causes Adipose Hypertrophy and Alterations in Metabolism of Complex Lipids under High Fat Diet. J. Biol. Chem. 286, 35578-35587 (2011

Species mouse

Application IHC (paraffin)

Publication Macpherson, R. E. K. et al. Subcellular localization of skeletal muscle lipid droplets and PLIN family proteins OXPAT and ADRP at rest and following contraction in rat soleus muscle. Am. J. Physiol. - Regul. Integr. Comp. Physiol. -(2011).

Species rat

Application IHC (frozen)

Publication Timmers, S. et al. Paradoxical Increase in TAG and DAG Content Parallel the Insulin Sensitizing Effect of Unilateral DGAT1 Overexpression in Rat Skeletal Muscle. PLoS One 6, (2011).

Species rat

Application WB

Publication Chang, B. H.-J., Li, L., Saha, P. & Chan, L. Absence of adipose differentiation related protein upregulates hepatic VLDL secretion, relieves hepatosteatosis, and improves whole body insulin resistance in leptin-deficient mice. J. Lipid Res. 51, 2132-42 (

Species mouse

Application WB,IHC (paraffin)

Publication Akhtar, N. et al. Molecular dissection of integrin signalling proteins in the control of mammary epithelial development and differentiation. Development 136, 1019-27 (2009).

Species mouse

Application WB,IHC (paraffin)

Publication Straub, B. K., Stoeffel, P., Heid, H., Zimbelmann, R. & Schirmacher, P. Differential pattern of lipid droplet-associated proteins and de novo perilipin expression in hepatocyte steatogenesis. Hepatology 47, 1936-1946 (2008).

Species human

Application WB,IHC

Publication Yamaguchi, T., Omatsu, N., Omukae, A. & Osumi, T. Analysis of interaction partners for perilipin and ADRP on lipid droplets*. Mol. Cell. Biochem. 284, 167-173 (2006).

Species mouse

Application ICC-IF

Publication Ohsaki, Y., Maeda, T. & Fujimoto, T. Fixation and permeabilization protocol is critical for the immunolabeling of lipid droplet proteins. Histochem. Cell Biol. 124, 445-452 (2005).

Species human

Application ICC-IF

Publication Fukumoto, S. & Fujimoto, T. Deformation of lipid droplets in fixed samples. Histochem. Cell Biol. 118, 423-428 (2002).

Species human

Application ICC-IF

Publication Heid, H. W., Moll, R., Schwetlick, I., Rackwitz, H. R. & Keenan, T. W. Adipophilin is a specific marker of lipid accumulation in diverse cell types and diseases. Cell Tissue Res. 294, 309-21 (1998).

Species human,mouse,rat,bovine,dog

Application WB,IHC (frozen),ICC-IF,IEM

Publication Heid, H. W., Lzer, M. S. & Keenan, T. W. Adipocyte differentiation-related protein is secreted into milk as a constituent of milk lipid globule membrane. Biochem. J. 320, 1025-1030 (1996).

Species bovine

Application WB

Downloads

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Filetype

File IHC guinea pig antibody

Category Protocol

Size 167.92 KB

Filetype pdf

File WB protocol

Category Protocol

Size 210.69 KB

Filetype pdf

Datasheet

MSDS

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.

The fixation method used for lipidic material influences tremendously the quality of staining results.
Tissue fixation should not exceed 2% paraformaldehyde.
Heat induced antigen retrieval is recommended. In many cases a standard protocol using 10 mM citrate buffer (pH 6) works fine.

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.

Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.

The use of milk preparations (skim milk, dry milk) for blocking and for antibody dilution buffer might be problematic. The antigen of the Perilipin 2 antibodies was originally found and described in bovine milk and 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. In this case Tween 20 or BSA or ovalbumin is recommended as e.g. blocking material.
Immune reaction with cultured cells: this depends on the age of the cell culture; old cells (more than confluent) tend to accumulate fat and are therefore positive for Perilipin 2. Freshly split cells contain only few lipid droplets and are not easily detectable as positive for Perilipin 2.
Tissue with a high contents of fat might be problematic in electrophoretical separation. One has to load much material onto the SDS gel in order to get enough protein for a positive immunological signal. In addition the protein solubility is sometimes hindered by high fat concentration in the sample.

Positive signals from e.g. cultured cells depend on the age of the cell culture: old cells (more than confluent) tend to accumulate fat and are therefore positive for Perilipin 2. Freshly split cells contain only few lipid droplets and are not easily detectable as positive for Perilipin 2. With old /aged cells, however, there is always a problem with potential degradation processes.

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.

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