Reagents needed for the detection of labeled interleukin 6 by acridinium ester chemiluminescence method
The chemiluminescence method in the antibody detection of the new coronavirus is one of the important detection methods. In addition to detecting various antibodies, the commonly used luminescent reagent acridinium ester can also be used for the label detection of a variety of biological macromolecules, such as important in the defense mechanism of the human body. Detection of the cytokine interleukin 6.
Interleukin 6 can be produced by cells with a variety of immune functions or other cells. Its main role is to promote the proliferation and differentiation of B cells and the secretion of antibodies. It also has a wide range of effects on liver cells, T cells, nervous tissues, hematopoietic system, and anti-tumor effects, etc. .
It can also be called beta 2 interferon, hepatocyte stimulating factor, hybridoma growth factor, B cell differentiation factor and so on.
Chemiluminescence reagent acridinium ester
Reagents needed for the detection of interleukin 6 with acridinium esters:
A highly sensitive and specific acridinium ester chemiluminescence method for the detection of interleukin 6. The kit includes sample diluent, magnetic particles coated with interleukin 6 monoclonal antibody, interleukin 6 monoclonal antibody labeled with chemiluminescence markers, Excitation solution A, excitation solution B, interleukin 6 calibrator solution and cleaning solution.
Acridine esters commonly used to label interleukin 6 include NSP-DMAE-NHS, NSP-SA-NHS and so on. The molar ratio of acridinium ester to antibody is 5-10:1, and the preferred molar ratio is 7.4:1.
The sample dilution buffer is a phosphate buffer with a pH of 7.4, containing 1% bovine serum albumin, 0.1% TritonX-100 and 0.3% Proclin.
The surface modification group of the magnetic beads used can be carboxyl, amino, streptavidin-biotin or p-toluenesulfonyl, and the particle size of the magnetic beads can be 1.5-3 μm. The magnetic particles can be directly coupled with interleukin 6 antibody, or the magnetic particles can be coupled with streptavidin, and biotin-labeled interleukin 6 antibody is used at the same time.
The interleukin-6 monoclonal antibody is a specific monoclonal antibody for the interleukin-6 to be tested.
The preparation process for the detection of interleukin 6 with acridinium ester:
1) Prepare sample dilution buffer.
2) Preparation of magnetic particles coated with interleukin-6 monoclonal antibody: magnetically separate the carboxylated magnetic particle solution, remove the supernatant, resuspend it in MES buffer, add EDC solution, activate it on a vertical rotator, and remove it after activation. The magnetic particles and the interleukin 6 antibody are coupled on a vertical rotator, the supernatant is removed by magnetic separation, washed 3 times, and then a buffer containing 1% BSA is added for blocking, and finally the magnetic particle suspension is placed at 2-8℃ save.
In addition to NSP-DMAE-NHS and NSP-SA-NHS, the two acridinium ester luminescent reagents, Desheng has also developed 4 acridinium esters to better match different antibodies, proteins, and nucleic acids waiting to be detected.
MOPS buffer is an important biochemical reagent used to maintain the acid-base balance and ion environment of tissue samples, protecting cell structure and function. Widely used in cell culture, tissue fixation, and immunohistochemical staining to improve experimental accuracy and reliability. Understanding the characteristics and application principles of MOPS buffer is crucial for biomedical research.