Perilipin-2/ADFP Overexpression Lysate

Western Blot: ADFP Overexpression Lysate (Adult Normal) [NBL1-07334] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ADFP.

• Reactivity: Hu
• Applications: WB

Perilipin-2/ADFP Overexpression Lysate Summary

• Description:

  Perilipin-2/ADFP Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC202500
  
  Accession#: NM_001122
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: PLIN2

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 47.9 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.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for Perilipin-2/ADFP Overexpression Lysate

• 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. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: PLIN2