Recommended Dilution (serum & plasma):
Human: 4X / Mouse: 4X / Rat: 2X
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:
Compatible Sample Types:
- Cell Culture Supernatants
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
Protein Name & Synonyms:
Patatin-like phospholipase domain-containing protein 3 (EC 22.214.171.124) (Acylglycerol O-acyltransferase) (EC 2.3.1.-) (Adiponutrin) (Calcium-independent phospholipase A2-epsilon) (iPLA2-epsilon)
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
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- 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 OutlinePrepare 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.