ENOS ELISA Kit

Catalog No. ABIN4886056

This Colorimetric ELISA kit is designed for the quantitative measurement of Human ENOS in Cell Lysate and Tissue Lysate.

Quick Overview for ENOS ELISA Kit (ABIN4886056)

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

Binding Specificity: pSer1177, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details ENOS ELISA Kit

Purpose: Human Phospho-eNOS (S1177) and Total eNOS ELISA Kit. This assay semi-quantitatively measures phophorylated eNOS (Ser1177) and Total eNOS in lysate samples.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes human eNOS phosphorylated at site Serine-1177as well as total eNOS

Characteristics:

• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-Phospho Antibody
• Anti-Pan Antibody
• HRP-Conjugated Secondary Antibody
• Streptavidin-Conjugated HRP
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Benchtop rocker or shaker
• Microplate reader capable of measuring absorbance at 450 nm

Application Details

Sample Volume: 100 μL

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 μL of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared 1X HRP-Streptavidin to each well.
7. Incubate 1 h at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Expiry Date: 6 months

Target Details for ENOS

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

Alternative Name: 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