TMB Two-Component Substrate solution(for Elisa)

Kit Components:

At present, enzyme immunoassay (EIA) technology has been widely used in quantitative or qualitative detection of antigens, haptens or antibodies. Horseradish peroxidase (HRP) and its conjugate are commonly used in enzyme-linked immunoassay (ELISA) because of 3,3’ 5, 5, & # 39; Tetramethylbenzidine (TMB) is widely used in the color reaction system of HRP because it has higher sensitivity and no carcinogenicity than other color gen. TMB is mainly used in enzyme-linked immunosorbent assay (ELISA), immune dot hybridization or immunohistochemistry, and chlorine and hydrogen peroxide detection analysis. In order to meet the development and production of different kit products and scientific research needs, the company has specially developed different types of TMB color developing solution for immunoanalysis based on HRP.

Appearance and structure: Color developing liquid A should be colorless or light yellow transparent liquid, color developing liquid B should be colorless transparent liquid. There were no precipitates, particles and flocculents.

Used in the chromogenic stage of enzyme-linked immunoassay: It can react on the sample to produce a dark blue product.

Usage:

Add liquid: After HRP binding is added to the sample hole and incubated for A certain period of time, wash the plate with appropriate washing solution 3-5 times, add 50ul of substrate developing solution A and B to each hole (A and B are mixed first and then added to the hole), gently mix well, and incubate at room temperature (15-25℃) or 37℃ for 10-30 minutes or longer according to personal experimental needs. Until the color is the desired shade.

Termination: Add 1M hydrochloric acid or sulfuric acid solution of equal volume to each hole to terminate the reaction, and the reaction liquid in the hole changes from blue to yellow.

Reading: The light absorption value of each pore solution was measured at 450nm within 15 minutes after termination of the reaction.

Control: The antibody/antigen of blank control without HRP labeling and the negative control results should be colorless. The positive control and HRP labeled antibody/antigen produced dark blue products.

Note: If a high reaction background or precipitation occurs, it indicates that the TMB substrate is reacting too strongly. In order to avoid precipitation, it can be read immediately after termination; Or further dilute the primary antibody and/or HRP conjugate.

Note:

1, the experimental supplies should be specific to avoid cross contamination.

2, TMB is irritating to the human body, please pay attention to appropriate protection.

3. For your safety and health, please wear a lab coat and disposable gloves

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.