Mouse Visfatin EasyTest™ EIA Kit

Product Features

Product Name           Mouse Visfatin EasyTest™ EIA Kit
Assay Type                   Solid Phase Competitive ELISA
Format                           96-well strip plate
Assay Process             Two wash step, three incubations
Assay Length              2.5 hours
Sample Volume         Serum , Plasma, 50µL
Detection Method    Colorimetric
Sensitivity                   130pg/mL
Assay Range                9.8-40000pg/mL
Reactivity                     Mouse
Cross-Reactivity       No cross-reactivity observed with available related molecules
Gene Names               NAMPT; 1110035O14Rik; AI314458; AI480535; NAmPRTase; Pbef; Pbef1; Visfatin
Gene ID                         59027
Swissprot ID               Q99KQ4

Standard Curve        Mouse Visfatin EasyTest™ EIA Kit

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Anti-Visfatin Antibody
• Standard Visfatin peptide
• Biotinylated Visfatin peptide
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 25 µl of anti-Visfatin antibody to all wells.
3. Add 75 µl of 1 x assay diluent to the blank wells and 50 µl to B0 wells.
4. Add 50 µl of standard peptide/samples to the appropriate wells.
5. Add 25 µl of biotinylated peptide into each well except the blank wells.
6. Incubate 1.5 h at RT.
7. Wash and add 100 µl of diluted streptavidin-HRP solution to all wells.
8. Incubate 45 min at RT.
9. Wash and add 100 µl of TMB Substrate Solution into each well.
10. Incubate 15 min at RT.
11. Add 50 µl of Stop Solution to each well.
12. Read at 450 nm/570 nm.

Background

Visfatin (also known as PBEF and Namp) is an enzyme that in humans is encoded by the PBEF1 gene. This protein has also been reported to be a cytokine (PBEF) that promotes B cell maturation and inhibits neutrophil apoptosis. It is downregulated by an increase of miR-34a in obesity via a 3’UTR functional binding site of NAMPT mRNA resulting in a reduction of NAD (+) and decreased SIRT1 activity.

Visfatin catalyzes the condensation of nicotinamide with 5-phosphoribosyl-1-pyrophosphate to yield nicotinamide mononucleotide, one step in the biosynthesis of nicotinamide adenine dinucleotide. The protein is an adipokine that is localized to the bloodstream and has various functions, including the promotion of vascular smooth muscle cell maturation and inhibition of neutrophil apoptosis. It also activates insulin receptor and has insulin-mimetic effects, lowering blood glucose and improving insulin sensitivity.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Mouse Visfatin EasyTest™ EIA Kit is for Research Use Only.

• Manual
• AssayProcedure
• MSDS