RayBio® Human, Mouse and Rat Phosphotyrosine iNOS ELISA Kit. This assay semi-quantitatively measures tyrosine-phosphorylated iNOS in lysate samples.PEL-iNOS-YPEL-iNOS-Y
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Species | Human, Mouse, Rat |
| Specificity | This ELISA kit recognizes human iNOS phosphotyrosine. |
| Compatible Sample Types | Tissue Lysates, Cell Lysates |
| Accession Number | P35228|P29477|Q06518 |
| Gene Id | 4843 |
| Gene Symbols | NOS2, NOS2A |
| Protein Name / Synonyms | Nitric oxide synthase, inducible (EC 1.14.13.39) (Hepatocyte NOS) (HEP-NOS) (Inducible NO synthase) (Inducible NOS) (iNOS) (NOS type II) (Peptidyl-cysteine S-nitrosylase NOS2) |
| Quantitative/Semi-Quantitative | Semi-Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Post-Translational Modifications, Phosphorylation, Tyrosine Kinase Signaling |
| Shipping Type | Blue ice |
| Storage | -20°C |
Product Features
- Rapidly measure phosphorylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Biotinylated Anti-Phosphotyrosine Antibody
- Stop Solution
- Assay Diluent(s)
- Positive Control Sample
- Lysis Buffer
- Streptavidin-Conjugated HRP
- TMB One-Step Substrate
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
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 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.
Typical Data
Positive Control
THP1 cells were treated with Pervanadate. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Pervanadate (PV) Stimulation of THP1 Cell Line
THP1 cells were untreated or treated with Pervanadate for 10 min at 37°C. Cell lysates were analyzed using this phosphoELISA:
Storage/Stability
Upon receipt, the kit should be stored at â20°C. Please use within 6 months from the date of shipment.
