ENOS ELISA Kit

Catalog No. ABIN8070273

This Colorimetric ELISA kit is designed for the quantitative measurement of Human ENOS.

Quick Overview for ENOS ELISA Kit (ABIN8070273)

Target: ENOS (NOS3) (Nitric Oxide Synthase 3 (Endothelial Cell) (NOS3))

Binding Specificity: citrullinated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details ENOS ELISA Kit

Purpose: Measures citrullinated eNOS (NOS3) and total eNOS in human cell and tissue lysates, enabling precise assessment of eNOS citrullination by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-eNOS (NOS3) Antibody
• Anti-Citrullination Detection Antibody
• Anti-eNOS (NOS3) Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• 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
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare sample dilutions
• Distilled or deionized water
• Shaker

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed.
2. Add 100 μL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Target Details for ENOS

Target: ENOS (NOS3) (Nitric Oxide Synthase 3 (Endothelial Cell) (NOS3))

Alternative Name: eNOS

Background:

Gene Symbols: NOS3

Background: The Citrullinated-eNOS (NOS3) and Total-eNOS (NOS3) ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human citrullinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan eNOS (NOS3) antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and eNOS (NOS3) present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-citrullination antibody is used to detect citrullinated eNOS (NOS3) or biotinylated anti-eNOS (NOS3) antibody is used to detect total eNOS (NOS3) . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of citrullinated eNOS (NOS3) or total eNOS (NOS3) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): Nitric oxide synthase 3, Constitutive NOS (cNOS), EC-NOS, NOS type III (NOSIII), Nitric oxide synthase, endothelial (Endothelial NOS, eNOS)

Gene ID: 4846

UniProt: P29474

Pathways: ACE Inhibitor Pathway, Regulation of Systemic Arterial Blood Pressure by Hormones, Cellular Response to Molecule of Bacterial Origin, Myometrial Relaxation and Contraction, Signaling Events mediated by VEGFR1 and VEGFR2, Thromboxane A2 Receptor Signaling, VEGFR1 Specific Signals, VEGF Signaling