Protein Name & Synonyms:Patatin-like phospholipase domain-containing protein 3 (EC 3.1.1.3) (Acylglycerol O-acyltransferase) (EC 2.3.1.-) (Adiponutrin) (Calcium-independent phospholipase A2-epsilon) (iPLA2-epsilon)
Detection Range:0.1-1,000 ng/ml
Recommended Dilution (serum & plasma):Human: 4X / Mouse: 4X / Rat: 2X
Specificity:This kit detects both long form (481aa) and short form (477aa) Adiponutrin proteins. No other active isoforms have been reported.Cross Reactivity: This EIA kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, Angiotensin II, NPY and APC.
Number of Targets Detected:1
Compatible Sample Types:- Cell Culture Supernatants
- Serum
Method of Detection:Colorimetric
Quantitative/Semi-Quantitative: Solid Support:96-well Microplate
Size:1, 2, or 5 x 96-Well Strip Microplate Kit
Human Adiponutrin (ADPN) is a 53 kD adipokine protein belong to the PNPLA family of phospholipase A2 enzymes. The 481 amino acid human Adiponutrin is a type II transmembrane protein, consisting of the patatin domain (aa 10 - 179) with enzyme activity and a transmembrane segment (aa 42 - 62). Human and mouse adiponutrins share 61% homology in amino acid sequence. Adiponutrin is mainly expressed in adipocytes and serves as a triacylglycerol lipase that mediates triacylglycerol hydrolysis in adipocytes. Reports have shown that Adiponutrin plays an important role in the balance of energy usage/storage in adipocytes. Its expression level is enhanced in response to insulin. Studies by Dr Clement"s group have shown that in obese subjects, short- or long-term calorie restriction diets were associated with a reduction in Adiponutrin gene expression. Adiponutrin mRNA level was previously shown to be negatively correlated with fasting glucose plasma levels and associated with insulin sensitivity of non-diabetic obese and non-obese subjects. Reports from the same group have also shown a strong relationship between Adiponutrin expression, insulin sensitivity and glucose metabolism in human adipose tissue.Other names for Adiponutrin include PNPLA3 (patatin-like phospholipase domain-containing protein 3).RayBio® Adiponutrin EIA kit targets the common sequence of human, mouse, and rat. This kit may be used to detect Adiponutrin expression in all these species with high specificity and sensitivity.
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- HRP-Streptavidin
- TMB One-Step Substrate
- Stop Solution
- Assay Diagram
- Positive Control Sample
- Capture Antibody
- User Manual
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Orbital shaker
- Aluminum foil
- Saran Wrap
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- SigmaPlot software (or other software that can perform four-parameter logistic regression models)
Protocol Outline
- Prepare all reagents, samples and standards as instructed.
- Add 100 µl detection antibody to each well.
- Incubate 1.5 h at RT or O/N at 4°C.
- Add 100 µl standard or sample to each well.
- Incubate 2.5 h at RT.
- Add 100 µl prepared streptavidin solution.
- Incubate 45 min at RT.
- Add 100 µl TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl Stop Solution to each well.
- Read plate at 450 nm immediately.
Standard, Biotinylated Adiponutrin peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.