Perilipin-2/ADFP Antibody - BSA Free

Simple Western: Perilipin-2/ADFP Antibody [NB110-40878] - 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.

Immunohistochemistry-Paraffin: Perilipin-2/ADFP Antibody [NB110-40878] - ADFP antibody was tested in mouse liver using DAB with hematoxylin counterstain.

Immunohistochemistry-Frozen: Perilipin-2/ADFP Antibody [NB110-40878] - Detection of PLIN2 in pig sleletal muscle cross section. Antibody dilution 1:100 in PBS with 2% normal goat serum. Secondary antibody: Alexa Fluor 488 goat anti-rabbit IgG at 1:1000. IHC-Fr image submitted by a verified customer review.

Western Blot: Perilipin-2/ADFP Antibody [NB110-40878] - Detection of human ADFP in human liver lysate.

Western Blot: Perilipin-2/ADFP Antibody [NB110-40878] - CDK5 and 14-3-3iu and KIAA0528 are found on lipid droplet (LD) cargo. Intact purified LDs were probed with antibodies against CDK5, 14-3-3iu, KIAA0528, and GFP (negative control), the percentage of LDs that had detectable immunofluorescence signal quantification is shown in a and sample immunofluorescence images are shown in b with 2 i1/4m scale bars, and white arrows designate lipid droplets. To assess CDK5 localization to LDs, proteins were precipitated from purified LDs and analyzed by WB. Perilipin-2: (an LD marker) is present on both samples and 25A- CDK5 is present on the LDs from the CDK5 overexpression background. Image collected and cropped by Citeab from the following publication (Regulation of in vivo dynein force production by CDK5 and 14-3-3u and KIAA0528. Nat Commun (2019) licensed under a CC-BY license.

Flow Cytometry: Perilipin-2/ADFP Antibody [NB110-40878] - An intracellular stain was performed on HepG2 cells with Perilipin-2 Antibody NB110-40878B (blue) and a matched isotype control (orange). Both antibodies were conjugated to Biotin. Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Streptavidin - R-Phycoerythrin Protein (2012-1000, Novus Biologicals).

Product Discontinued

Perilipin-2/ADFP Antibody - BSA Free Summary

• Immunogen: A synthetic peptide made to an internal portion of human ADFP (within residues 150-250) [Swiss-Prot# Q99541]
• Localization: Membrane, cytoplasm
• Predicted Species: Primate (99%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: PLIN2
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 2.5 ug/mL
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200
  • Simple Western 1:50
  • Western Blot 2 ug/ml
• Application Notes: This ADFP antibody is useful for Western blot analysis where a band is observed ~48 kDa, and Immunohistochemistry of paraffin embedded sections where cytoplasmic and membrane staining was observed in mouse liver. 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.
  
  This Perilipin-2/ADFP Antibody is validated for IHC-Fr from a verified customer review.
• Control:

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

• Control Peptide:

  Perilipin-2/ADFP Protein (NB110-40878PEP)

Reactivity Notes

Monkey reactivity reported in scientific literature (PMID: 30651536). Immunogen has 89% identity to porcine and 87% identity to bovine. Porcine reactivity reported from a verified customer review.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS, 30% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1.23 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 (NB110-40878)(5)

Publications using NB110-40878

• Al-Rashed F, Haddad D, Al Madhoun A et al. ACSL1 is a key regulator of inflammatory and macrophage foaming induced by short-term palmitate exposure or acute high-fat feeding iScience 2023-07-01 [PMID: 37416456] (WB, Human)
• Chapman DE, Reddy BJN, Huy B et al. Regulation of in vivo dynein force production by CDK5 and 14-3-3E and KIAA0528. Nat Commun 2019-01-16 [PMID: 30651536] (WB, Monkey)
• Fujisawa N, Yoshioka W, Yanagisawa H, Tohyama C. Roles of cytosolic phospholipase A2a in reproductive and systemic toxicities in 2,3,7,8-tetrachlorodibenzo-p-dioxin-exposed mice. Arch. Toxicol. 2017-10-17 [PMID: 29043426] (WB, Mouse)
• Goh VJ, Tan JS, Tan BC et al. Postnatal deletion of Fat storage-inducing Transmembrane Protein 2 (FIT2/FITM2) causes lethal enteropathy. J. Biol. Chem. 2015-08-24 [PMID: 26304121] (IHC-P, WB, Mouse)
  
  Details:
  Perilipin-2/PLIN2 antibody used for WB and IHC-P on mouse enterocytes (gavaged vs. non-oil gavaged L/L and iF2KO mice at day 5 post-tamoxifen treatment, Figure 6).
• Dahl N Development of skeletal muscle and adipose tissues in neonatal dairy calves upon a maternal supplementation with essential fatty acids and conjugated linoleic acids Thesis 2022-01-01

Reviews for Perilipin-2/ADFP Antibody (NB110-40878) (3)

Average Rating: 3.3

(Based on 3 reviews)

We have 3 reviews tested in 3 species: Mouse, Bovine and Mouse, Pig.

reviewed by: Armen Petrosyan IHC-P Mouse 06/27/2023

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: Mouse Liver Tissue
• Species: Mouse
• Lot: D-6

reviewed by: Verified Customer IHC-Fr Pig 09/15/2020

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Skeletal muscle
• Species: Pig
• Lot: D-1

Comments

• Comments: no fluorescence was detected when blocking the antibody binding in advance with blocking peptide

reviewed by: Heather Talbott WB Bovine and Mouse 03/27/2017

Summary

• Application: Western Blot
• Sample Tested: bovine corpus luteum,bovine fat,bovine heart,bovine liver,Mouse heart,mouse liver
• Species: Bovine and Mouse
• Lot: C-1

Comments

• Comments: 20ug of sample/well
  Block - 30min at room temperature 5% non-fat dry milk in 0.1% Tween 20 in TBS
  Primary - 1:1000, 2h at room temperature 1% non-fat dry milk in 0.1% Tween 20 in TBS
  Secondary - 1:10,000 anti-rabbit-HRP [Jackson 111-035-003 Lot 122107], 1:5,000 anti-biotin-HRP [Cell Signaling 7075P Lot 28] 2h at room temperature 1% non-fat dry milk in 0.1% Tween 20 in TBS
  ECL - SuperSignal Fempto
  UVP Biospectrum

FAQs for Perilipin-2/ADFP Antibody (NB110-40878). (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. May I ask what the recommended blocking buffer is for NB110-40878?
   • We validated this antibody using 5% milk plus 1% BSA in TBS-T as our blocking buffer for Western blot. This is our standard blocking buffer it isn't listed in the product specific protocol.

Bioinformatics

• Gene Symbol: PLIN2