Human Adiponutrin/PNPLA3 Antibody

Adiponutrin/ADPN in Human Adipocytes.

Adiponutrin/ADPN was detected in immersion fixed human adipocytes using 10 µg/mL Sheep Anti-Human Adiponutrin/ADPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5208) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counter-stained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Adiponutrin/ADPN by Western Blot.

Western blot shows lysates of human adipose tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Adiponutrin/ADPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5208) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Adiponutrin/ADPN at approximately 53 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Adiponutrin/PNPLA3 by Immunocytochemistry/ Immunofluorescence

ABHD5 interacts with multiple PNPLA family members.A, schematic of PNPLA4 fusions. B, fluorescent imaging of U2OS cells transfected with YFP-tagged PNPLA4, PNPLA3 WT, PNPLA3-PNPLA4 fusion, or PNPLA3-PNPLA4 fusionAAEE mutant to show lipid droplet targeting. Cells were treated with 0.2 mM oleic acid overnight and stained with Mitotracker Red and Lipi-Blue. Images are representative of results seen in three technical replicates of three experiments. C, fluorescent imaging and FRET analysis of COS-7 cells transfected with YFP-tagged PNPLA4, PNPLA3 WT, or PNPLA3 mutant and ECFP-tagged ABHD5 R229N. Cells were treated with 0.2 mM oleic acid overnight and stained with LipidTOX Neutral Red. Images are representative of results seen in three technical replicates of three consecutive experiments. D, Gluc protein complementation assay of HEK293A cells transfected with various PNPLA family proteins tagged with GlucN and ABHD5-GlucC. Cells were treated with 0.2 mM oleic acid in 10% BSA or 10% BSA alone overnight. Data show that oleic acid treatment significantly alters the interaction of ABHD5 with PNPLA3 WT, PNPLA3 I148M, and PNPLA5. Data represent averages of three trials with four technical replicates per trial. Statistics calculated using two-way ANOVA with Sidak’s multiple comparison test, ∗p < 0.05. The scale bar represents 10 μm. ABHD5, alpha/ beta hydrolase domain-containing protein 5; BSA, bovine serum albumin; PNPLA, patatin-like phospholipase domain. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39814233), licensed under a CC-BY license. Not internally tested by R&D Systems.