The RayBiotech Protein Carbamylation Quantitative ELISA is a very rapid, convenient, and sensitive assay kit that can monitor the total levels of protein carbamylation in cell and tissue lysates.
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Compatible Sample Types | Tissue Lysates, Cell Lysates |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Shipping Type | Blue ice |
| Storage | -20°C |
Introduction
RayBio® Protein Carbamylation Quantitative ELISA kit is a very rapid, convenient, and sensitive assay kit that can monitor the total levels of protein carbamylation in tissue and cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of carbamylated proteins. An anti-pan carbamylation antibody has been coated onto a 96-well plate. Standard and samples are pipetted into the wells and carbamylated protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-carbamylation antibody is used to detect only carbamylated protein. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted to the wells. The wells are washed again, a TMB substrate solution is added to the wells and color develops in proportion to the amount of carbamylated protein bound. The stop solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Product Features
- Rapidly quantitate protein carbamylation
- Screen numerous different 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 Anti-Carbamylation Microplate
- Wash Buffer
- Carbamylated Protein Standard
- Anti-Carbamylation Antibody
- HRP-conjugated anti-rabbit IgG concentrate
- TMB One-Step Substrate Reagent
- Stop Solution
- Assay Diluent
- Lysis Buffer
Other Materials Required
- Microplate reader capable of measuring absorbance at 450 nm
- Protease and Phosphatase inhibitors
- Shaker
- 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
- Absorbent paper
- Distilled or deionized water
- Log-log graph paper or computer and software for ELISA data analysis
- Tubes to prepare the standard or sample dilutions
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 each standard and sample to each well. Incubate 2.5 hours at room temperature or overnight at 4°C with gentle shaking.
- Add 100 μl prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
- Add 100 μl prepared HRP-Conjugated solution. Incubate for 1 hour at room temperature with gentle shaking.
- Add 100 μl TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
- Add 50 μl Stop Solution to each well. Read at 450 nm immediately.
Typical Data
Standard Curve
Storage/Stability
The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles. For extended storage, it is recommended to store at -80°C. For prepared reagent storage, see manual.