Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

RayBio® Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit. This assay semi-quantitatively measures eNOS phosphorylated at Serine-1177 as well as total eNOS in adherent cell lines.

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

RayBio® Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit. This assay semi-quantitatively measures eNOS phosphorylated at Serine-1177 as well as total eNOS in adherent cell lines.

Product Description

Specifications

Size	1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Estimated Lead Time	4-6 weeks
Species	Human
Specificity	This ELISA kit recognizes Human eNOS phosphorylated at site Serine-1177 as well as total eNOS.
Compatible Sample Types	Adherent Cell Culture
Accession Number	P29474
Gene Id	4846
Gene Symbols	NOS3
Protein Name / Synonyms	Nitric oxide synthase, endothelial (EC 1.14.13.39) (Constitutive NOS) (cNOS) (EC-NOS) (Endothelial NOS) (eNOS) (NOS type III) (NOSIII)
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Design Principle	Cell-based
Method Of Detection	Colorimetric
Research Area	Angiogenesis, Cardiovascular Disease, Post-Translational Modifications, Phosphorylation, HIF-1 alpha Signaling, PI3K-AKT Signaling
Shipping Type	Blue ice
Storage	-20°C

Product Features

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

Application Notes

Kit 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

Other Materials Required

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

Protocol Outline

Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacturer's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37°C.
Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Storage/Stability

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.

Have this product in hand?

Get your lot-specific datasheet at Ray's Docs. The datasheet linked below is our current version and may not match your lot.

Datasheet (CBEL-prelim.pdf)

SDS (ec571c3342d1422f27802ee7dc52b894dbb8f17c.pdf)

Product Features

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

Application Notes

Kit 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

Other Materials Required

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

Protocol Outline

Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacturer's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37°C.
Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Storage/Stability

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.

Featured Products

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Product Description

Specifications

Product Features

Application Notes

Storage/Stability

Have this product in hand?

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Product Description

Specifications

Product Features

Application Notes

Storage/Stability

Have this product in hand?

Featured Products

Featured Products

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Product Description

Specifications

Product Features

Application Notes

Storage/Stability

Documents

Have this product in hand?

Reviews

Ask A Question

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Custom Human Phospho-eNOS (Ser1177) and Total eNOS Cell-Based ELISA Kit

Product Description

Specifications

Product Features

Application Notes

Storage/Stability

Documents

Have this product in hand?

Reviews

Ask A Question

Featured Products