Perilipin-2/ADFP Antibody - BSA Free

Dual RNAscope ISH-IHC: Perilipin-2/ADFP Antibody [NB110-40877] - Formalin-fixed paraffin-embedded tissue sections of mouse intestine were probed for Perilipin-2/ADFP mRNA (ACD RNAScope Probe, catalog #577111; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Rabbit Polyclonal (Novus Biologicals catalog # NB110-40877) at 1:150 dilution with overnight incubation at 4 degrees Celsius followed by incubation with anti-rabbit IgG VisUCyte HRP Polymer Antibody (Catalog # VC003) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to intestinal villi.

Simple Western: Perilipin-2/ADFP Antibody [NB110-40877] - Simple Western lane view shows a specific band for ADFP in 0.5 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: Perilipin-2/ADFP Antibody [NB110-40877] - Oleic acid increasesI2 oxidation, glucose utilization and expression of genes involved in LCFA metabolism in U138 GBM cells. Cropped western blot images and quantitation of PLIN2 and GAPDH protein levels from cell lysates. Human hepatoma (VL17A) cell line was used as a positive control (+control). Full length blots are in Supplementary Fig. A1. N=3 independent experiments in triplicate. Image collected and cropped by Citeab from the following publication (Lipid accumulation and oxidation in glioblastoma multiforme. Sci Rep (2019)) licensed under a CC-BY licence.

Immunohistochemistry: Perilipin-2/ADFP Antibody [NB110-40877] - ADFP antibody was tested in mouse liver using DAB with hematoxylin counterstain.

Immunocytochemistry/Immunofluorescence: Perilipin-2/ADFP Antibody [NB110-40877] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Perilipin-2/ADFP Antibody NB110-40877 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Western Blot: Perilipin-2/ADFP Antibody [NB110-40877] - Detection of murine ADFP in mouse liver lysate.

Immunocytochemistry/Immunofluorescence: Perilipin-2/ADFP Antibody [NB110-40877] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-Perilipin-2/ADFP at a 1:100 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: Perilipin-2/ADFP Antibody [NB110-40877] - HepG2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-Perilipin-2/ADFP Antibody NB110-40877 at 1 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

• Reactivity: Hu, Mu, Rt, Bv
• Applications: WB, Simple Western, ICC/IF, IHC, IHC-Fr, IHC-P, Dual ISH-IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Perilipin-2/ADFP Antibody - BSA Free Summary

• Immunogen: A synthetic peptide made to a C-terminal region of mouse ADFP (within residues 350-425) [Swiss-Prot# P43883]
• Localization: Membrane
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PLIN2
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Dual RNAscope ISH-IHC
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200
  • Simple Western 1:50
  • Western Blot 2 ug/ml
• Application Notes: Use in Immunohistochemistry-Frozen reported in scientific literature (PMID:35041644) This ADFP antibody can be used for Western blot and Immunofluorescence/Immunocytochemistry. In Western blot a band is observed at approx. 51 kDa. In ICC/IF, membrane staining was observed in HeLa cells. In IHC-P, staining was observed on the membrane of mouse liver cells Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 51 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  Human Adrenal Whole Tissue Lysate (Adult Whole Liver Cirrhosis)
  Human Liver Whole Tissue Lysate (Adult Whole Normal)
  Mouse Liver Whole Tissue Lysate (Adult Whole Normal)
  Perilipin-2/ADFP Overexpression Lysate

• Control Peptide:

  Perilipin-2/ADFP Protein (NB110-40877PEP)

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 29053516). Use in Bovine reported in scientific literature (PMID:32647143).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Perilipin-2/ADFP Antibody - BSA Free

• ADFP
• Adipophilin
• Adipose differentiation-related proteinadipophilin
• ADRP
• MGC10598
• perilipin 2
• Perilipin2
• Perilipin-2
• PLIN2

Background

Perilipin-2/Plin-2, also known as adipose differentiation-related protein (ADFP), ADRP, and adipophilin, is a member of the perilipin family of lipid droplet (LD) proteins that functions in lipid accumulation, lipid storage, and LD formation (1,2). Perilipin-2 is ubiquitously expressed and is the major LD protein of the liver (1-3). Human perilipin-2 is 437 amino acids (aa) in length with a theoretical molecular weight of 48 kDa (1,4). Human and mouse perilipin-2 share 81.8% sequence identity. One defining structural feature of perilipin-2 and other LD-related proteins is formation of amphipathic alpha helices (1,3,5). The perilipin-2 protein contains a PAT (perilipin A, ADRP, TIP47) domain, a 11-mer repeat motif, and a 4-helix bundle (1,5). The PAT domain has a role in triglyceride (TG) stabilization, the 11-mer repeat motif functions in cytoplasmic LD binding, and the 4-helix bundle facilitates membrane binding (5). Unlike LDs coated with other perilipin family members (e.g., perilipin-1 and perilipin-5), LDs coated with perilipin-2 are more permissive to lipolysis and protective of lipids and TGs (1-3). Studies have revealed that perilipin-2 knockout (KO) in mice is associated with resistance to fatty liver disease and diet-induced obesity (1). On the other hand, overexpression of perilipin-2 causes increased lipid and TG accumulation, which is a feature of many metabolic, cardiovascular, and age-related diseases (2). Additional research has found higher levels of perilipin-2 is associated with certain cancers such as colorectal cancer and renal carcinomas, and it may be a useful biomarker for detection (2). Though more research is needed, it is possible that inhibition or blocking of perilipin-2 may be a key strategy to combatting several pathologies (2).

References

1. Itabe H, Yamaguchi T, Nimura S, Sasabe N. Perilipins: a diversity of intracellular lipid droplet proteins. Lipids Health Dis. 2017;16(1):83. https://doi.org/10.1186/s12944-017-0473-y

2. Conte M, Franceschi C, Sandri M, Salvioli S. Perilipin 2 and Age-Related Metabolic Diseases: A New Perspective. Trends Endocrinol Metab. 2016;27(12):893-903. https://doi.org/10.1016/j.tem.2016.09.001

3. Sztalryd C, Brasaemle DL. The perilipin family of lipid droplet proteins: Gatekeepers of intracellular lipolysis. Biochim Biophys Acta Mol Cell Biol Lipids. 2017;1862(10 Pt B):1221-1232. https://doi.org/10.1016/j.bbalip.2017.07.009

4. Uniprot (Q99541)

5. Chong BM, Reigan P, Mayle-Combs KD, Orlicky DJ, McManaman JL. Determinants of adipophilin function in milk lipid formation and secretion. Trends Endocrinol Metab. 2011;22(6):211-217. https://10.1016/j.tem.2011.04.003

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Perilipin-2/ADFP Antibody (NB110-40877)(49)

Publications using NB110-40877

• Ibayashi M, Aizawa R, Mitsui J, Tsukamoto S Lipid droplet synthesis is associated with angiogenesis in mouse ovarian follicles Biology of reproduction 2022-12-28 [PMID: 36579469]
• Wang N, Wang M, Jeevaratnam S et al. Opposing effects of apoE2 and apoE4 on microglial activation and lipid metabolism in response to demyelination Molecular neurodegeneration 2022-11-23 [PMID: 36419137] (IF/IHC, Mouse)
• Lin M, Ku AT, Dong J et al. STAT5 confers lactogenic properties in breast tumorigenesis and restricts metastatic potential Oncogene 2022-10-19 [PMID: 36261627] (IF/IHC, Mouse)
• Fraunhoffer NA, Abuelafia AM, Bigonnet M et al. Multi-omics data integration and modeling unravels new mechanisms for pancreatic cancer and improves prognostic prediction NPJ precision oncology 2022-08-17 [PMID: 35978026] (IHC-P, Human)
  
  Details:
  Dilutions: 1:100
• Liu Q, Xiao JJ, Wang S et al. Paraoxonase 1 Ameliorates Renal Lipotoxicity by Activating Lipophagy and Inhibiting Pyroptosis The American journal of pathology 2022-08-10 [PMID: 35963464]
• Goossens P, Lu C, Cao J et al. Integrating multiplex immunofluorescent and mass spectrometry imaging to map myeloid heterogeneity in its metabolic and cellular context Cell metabolism 2022-07-12 [PMID: 35858629] (IHC-Fr, Mouse)
• Wang Z, Wu Q, Wang H et al. Diosgenin protects against podocyte injury in early phase of diabetic nephropathy through regulating SIRT6 Phytomedicine : international journal of phytotherapy and phytopharmacology 2022-06-13 [PMID: 35728388] (IF/IHC, Mouse)
• Antony R, Aby K, Gao H et al. UCHL1 Regulates Lipid and Perilipin 2 Level in Skeletal Muscle Frontiers in physiology 2022-04-07 [PMID: 35464088] (WB, Mouse)
• Asif S, Kim RY, Fatica T et al. Hmgcs2-mediated ketogenesis modulates high-fat diet-induced hepatosteatosis Molecular metabolism 2022-04-12 [PMID: 35421611] (IHC-P, Mouse)
• Lee W, Kim HY, Choi YJ et al. SNX10-mediated degradation of LAMP2A by NSAIDs inhibits chaperone-mediated autophagy and induces hepatic lipid accumulation Theranostics 2022-02-21 [PMID: 35265214] (WB, ICC/IF, Human, Mouse)
• Yu S, Wang L, Che D et al. Targeting CRABP-II overcomes pancreatic cancer drug resistance by reversing lipid raft cholesterol accumulation and AKT survival signaling Journal of experimental & clinical cancer research : CR 2022-03-08 [PMID: 35260193] (IF/IHC, Human)
• Lu Y, Zhou T, Xu C et al. Occludin is a target of Src kinase and promotes lipid secretion by binding to BTN1a1 and XOR PLoS biology 2022-01-01 [PMID: 35041644] (IHC-Fr, Mouse)
• Swinkels D, Das Y, Kocherlakota S et al. Cell Type-Selective Loss of Peroxisomal beta-Oxidation Impairs Bipolar Cell but Not Photoreceptor Survival in the Retina Cells 2022-01-04 [PMID: 35011723] (IF/IHC, WB, Mouse)
• Sorg O, Nocera T, Fontao F Et al. Lipid Droplets Proteins in Acne Skin. A sound target for the maintenance of low comedogenic sebum and acne-prone skin health. JID Innov 2021-12-15 [PMID: 34909752]
• Liu H, Wang C, Chang Z Et al. Losartan Attenuates Insulin Resistance and Regulates Browning Phenomenon of White Adipose Tissue in ob/ob Mice Curr Issues Mol Biol 2021-12-10 [PMID: 34889901]
• Rajesh Y, Reghupaty SC, Mendoza RG Et al. Dissecting the Balance Between Metabolic and Oncogenic Functions of Astrocyte-Elevated Gene-1/Metadherin Hepatology communications 2021-11-05 [PMID: 34741448] (IF/IHC, Mouse)
• Chen Q, Fang W, Cui K et al. Endoplasmic reticulum stress induces hepatic steatosis by transcriptional upregulating lipid droplet protein perilipin2 FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021-10-01 [PMID: 34547130]
• Xu S, Lee E, Sun Z et al. Perilipin 2 Impacts Acute Kidney Injury via Regulation of PPAR alpha Journal of immunology research 2021-09-09 [PMID: 34541006] (WB, Mouse)
• Casado ME, Huerta L, Marcos-DIaz A Et al. Hormone-sensitive lipase deficiency affects the expression of SR-BI, LDLr, and ABCA1 receptors/transporters involved in cellular cholesterol uptake and efflux and disturbs fertility in mouse testis Biochimica et biophysica acta. Molecular and cell biology of lipids 2021-08-27 [PMID: 34461308]
• Qi G, Mi Y, Shi X et al. ApoE4 Impairs Neuron-Astrocyte Coupling of Fatty Acid Metabolism Cell 2021-01-05 [PMID: 33406436] (Human)
• Lee LL, Khakoo AY, Chintalgattu V Cardiac pericytes function as key vasoactive cells to regulate homeostasis and disease FEBS Open Bio 2020-11-01 [PMID: 33135334] (WB, Mouse)
  
  Details:
  C57/BL6J mouse coronary smooth muscle cells underwent Western blot analysis.
• Talbott HA, Plewes MR, Krause C et al. Formation and characterization of lipid droplets of the bovine corpus luteum Sci Rep 2020-07-09 [PMID: 32647143] (WB, Bovine)
• Mayagoitia K, Shin SD, Rubini M et al. Short-term exposure to dietary cholesterol is associated with downregulation of interleukin-15, reduced thigmotaxis and memory impairment in mice Behav. Brain Res. 2020-06-22 [PMID: 32585301] (IF/IHC, Mouse)
• Chen L, Ma WL, Cheng WC et al. Targeting lipid droplet lysophosphatidylcholine for cisplatin chemotherapy J. Cell. Mol. Med. 2020-06-16 [PMID: 32543783] (WB, Human)
• Milutinovic DV, Brkljacic J, Teofilovic A et al. Chronic Stress Potentiates High Fructose-Induced Lipogenesis in Rat Liver and Kidney Mol Nutr Food Res 2020-05-07 [PMID: 32379936] (Rat)
• TaIb B, Aboussalah AM, Moniruzzaman M et al. Lipid accumulation and oxidation in glioblastoma multiforme Sci Rep 2019-12-20 [PMID: 31863022] (WB, Human)
• Helsley RN, Varadharajan V, Brown AL et al. Obesity-linked suppression of membrane-bound O-acyltransferase 7 (MBOAT7) drives non-alcoholic fatty liver disease Elife 2019-10-17 [PMID: 31621579] (Mouse)
• Asimakopoulou A, Vucur M, Luedde T et al. Perilipin 5 and Lipocalin 2 Expression in Hepatocellular Carcinoma Cancers (Basel) 2019-03-19 [PMID: 30893876] (WB, Human, Mouse)
• Farmer BC, Kluemper J, Johnson LA. Apolipoprotein E4 Alters Astrocyte Fatty Acid Metabolism and Lipid Droplet Formation Cells 2019-02-20 [PMID: 30791549] (WB)
• Chali F, Milior G, Marty S et al. Lipid markers and related transcripts during excitotoxic neurodegeneration in kainate-treated mice Eur. J. Neurosci. 2019-02-14 [PMID: 30767299] (IF/IHC, Mouse)
• Zhang P, Csaki LS, Ronquillo E et al. Lipin 2/3 phosphatidic acid phosphatases maintain phospholipid homeostasis to regulate chylomicron synthesis J. Clin. Invest. 2019-01-02 [PMID: 30507612] (WB, Human)
• Larsen Steen, Vigelso Andreas, Dandanell Sune et al. Simvastatin-Induced Insulin Resistance May Be Linked to Decreased Lipid Uptake and Lipid Synthesis in Human Skeletal Muscle: the LIFESTAT Study. J Diabetes Res. 2018-09-12 [PMID: 30276217] (WB, Human)
• Li Z, Rasmussen ML, Li J et al. Periodized low protein-high carbohydrate diet confers potent, but transient, metabolic improvements. Mol Metab 2018-08-28 [PMID: 30193785] (WB, Mouse)
• Li Z, Rasmussen M L et al. Low- and high-protein diets do not alter ex vivo insulin action in skeletal muscle. Physiol Rep 2018-01-07 [PMID: 29998629] (WB, Mouse)
• Madushani Herath KHIN, Cho J, Kim A et al. Phenolic acid and flavonoid-rich fraction of Sasa quelpaertensis Nakai leaves prevent alcohol induced fatty liver through AMPK activation J Ethnopharmacol. [PMID: 29906537] (IF/IHC, Mouse)
• Blossom SJ, Fernandes L, Bai S et al. Opposing actions of developmental trichloroethylene and high-fat-diet co-exposure on markers of lipogenesis and inflammation in autoimmune-prone mice. Toxicol. Sci. 2018-04-12 [PMID: 29669109] (IF/IHC, Mouse)
• Clemente C, Rius C, Alonso-Herranz L et al. MT4-MMP deficiency increases patrolling monocyte recruitment to early lesions and accelerates atherosclerosis. Nat Commun. 2018-03-02 [PMID: 29500407] (IHC-P, Mouse)
• Covington JD, Johannsen DL, Coen PM et al. Intramyocellular Lipid Droplet Size Rather Than Total Lipid Content is Related to Insulin Sensitivity After 8 Weeks of Overfeeding Obesity (Silver Spring). 2017-12-01 [PMID: 29071793] (Human)
• Kim SE, Lee JH, Kim TG et al. Fat Graft Survival After Recipient Site Pretreatment With Fractional Carbon Dioxide Laser Ann Plast Surg. 2017-12-01 [PMID: 29053516] (Rat)
• Li J, Mihalcioiu M, Li L et al. Vitamin D prevents lipid accumulation in murine muscle through regulation of PPARg and perilipin-2 expression. J. Steroid Biochem. Mol. Biol. 2017-10-14 [PMID: 29037825] (Mouse)
• Rendina-Ruedy E, Guntur AR, Rosen CJ et al. Intracellular lipid droplets support osteoblast function Adipocyte. 2017-07-19 [PMID: 28792783] (WB, Mouse)
• Vigelso A, Gram M, Wiuff C et al. Effects of immobilization and aerobic training on proteins related to intramuscular substrate storage and metabolism in young and older men. Eur. J. Appl. Physiol. 2015-12-01 [PMID: 26626913] (WB, Human)
• Nishikawa K, Iwaya K, Kinoshita M et al. Resveratrol increases CD68(+) Kupffer cells co-localized with adipose differentiation-related protein (ADFP) and ameliorates high-fat-diet-induced fatty liver in mice Mol Nutr Food Res. 2015-02-12 [PMID: 25677089] (WB, Mouse)
  
  Details:
  Perilipin-2/ADFP antibody (0.5 ug/ml) used for WB on mouse liver extract (Fig. 4).
• Bindesboll C, Berg O, Arntsen B et al. Fatty acids regulate perilipin5 in muscle by activating PPARdelta. J Lipid Res 2013-07-01 [PMID: 23606724]
• Lige B, Sampels V, Coppens I. Characterization of a second sterol-esterifying enzyme in Toxoplasma highlights the importance of cholesterol storage pathways for the parasite. Mol Microbiol 2013-02-03 [PMID: 23374239] (ICC/IF, Mouse)
• Chikka MR, McCabe DD, Tyra HM, Rutkowski DT. C/EBP Homologous Protein (CHOP) Contributes to Suppression of Metabolic Genes During ER Stress in the Liver. J Biol Chem 2012-12-31 [PMID: 23281479] (IHC-P, Mouse)
• Son S-H, Goo Y-H, Chang BH, Paul A. Perilipin 2 (PLIN2)-Deficiency Does Not Increase Cholesterol-Induced Toxicity in Macrophages PLoS ONE 2012-01-01 [PMID: 22427949] (WB, Mouse)
• Wohlers LM, Jackson KC, Spangenburg EE. Lipolytic signaling in response to acute exercise is altered in female mice following ovariectomy. J Cell Biochem. 2011-08-03 [PMID: 21815195]

Review for Perilipin-2/ADFP Antibody (NB110-40877) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Rat.

reviewed by: Verified Customer WB Rat 06/16/2020

Summary

• Application: Western Blot
• Sample Tested: Liver whole cell extract,Liver whole tissue homogenate, Sample Amount: 50ug
• Species: Rat
• Lot: F-3

Comments

• Comments: Concentration 1:1000, over night
  Secondary antibody: 1: 20000

FAQs for Perilipin-2/ADFP Antibody (NB110-40877). (Showing 1 - 2 of 2 FAQs).

1. I would appreciate if you are able to provide the exact amino acid sequence of the immunogens. The customer would like to check the percentage alignment with the chicken protein to determine which of the antiodies woudl be better to try in this species.
   • I am unable to provide you with this information however I can tell you that there is a 71% homology between the chicken protein and human protein. Our lab only recommends use if the homology is 85% or above.
2. I went trought your product datasheet, but I didn't find any information about the dilution I have to use for the western blot and if it's in 5% milk-TBST or other blocking buffer. Could you please provide me those information?
   • For our product NB110-40877, we recommend a starting dilution of 2 ug/ml for western blot. Our recommended diluent buffer is 5% nonfat milk in TBST.

Bioinformatics

• Gene Symbol: PLIN2
• Entrez:
  • 123(Human)
  • 11520(Mouse)
• Uniprot:
  • Q99541(Human)
  • P43883(Mouse)