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Perilipin-2/ADFP Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-07334

Novus Biologicals, part of Bio-Techne
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NBL1-07334

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Perilipin-2/ADFP Overexpression Lysate

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.

Product Specifications

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.

TMW

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.

Type

Overexpression

Scientific Data Images for Perilipin-2/ADFP Overexpression Lysate

Western Blot: Perilipin-2/ADFP Overexpression Lysate [NBL1-07334]

Western Blot: Perilipin-2/ADFP Overexpression Lysate [NBL1-07334]

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.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Perilipin-2

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

Alternate Names

ADFP, Adipophilin, Perilipin2, PLIN2

Gene Symbol

PLIN2

Additional Perilipin-2 Products

Product Documents for Perilipin-2/ADFP Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Perilipin-2/ADFP Overexpression Lysate

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.

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.

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