ENOS ELISA Kit

Catalog No. ABIN5526743

Product Details ENOS ELISA Kit

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

Binding Specificity: pSer1177, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Purpose: Human Phospho-CDK1 (Thr161) and Total CDK1 Cell-Based ELISA Cell-Based ELISA Kit. This assay semi-quantitatively measures eNOS phosphorylated at Serine-1177 as well as total eNOS in adherent cell lines.

Sample Type: Adherent Cell Culture

Analytical Method: Semi-Quantitative

Specificity: This ELISA kit recognizes Human eNOS phosphorylated at site Serine-1177 as well as total eNOS.

Characteristics:

• Rapidly measure phosphorylated protein in adherent cell lines
• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• No sample lysis is needed
• Compatible with a standard ELISA plate reader

Components:

• Uncoated 96-well Strip Microplate
• Wash Buffers
• Fixing Solution
• Quenching Buffer
• Blocking Buffer
• Anti-phospho antibody
• Anti-pan antibody
• HRP-Conjugated Secondary Antibody
• TMB One-Step Substrate
• Stop Solution

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

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

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 (NOS3 Products)

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