Plant hormone Elisa kit technical content - Database & Sql Blog Articles

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Kaixin micro test

Test probe P100-M3

EL-C1600N100013-B

The phytohormone elisa kit supplied by Shanghai Jinma Bio is currently in stock, and the order can be given priority shipping rights. Our company mainly quantifies the plant hormones (RADA) by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). The latter type is often used because of the lack of safety in the former. In ELISA, the detection of antigen-antibody reaction is achieved by means of an enzyme label, and the commonly used enzymes are horseradish peroxidase and alkaline phosphatase. The enzyme can directly label the hormone molecule, which is called the enzyme standard plant hormone, and can also be labeled with the second antibody (the antibody recognizing the anti-hormone antibody Fc fragment or the S. aureus A protein), which is called the enzyme-labeled secondary antibody. These two types of markers are used for solid phase antibody type and solid phase antigen type ELISA, respectively. A. Solid phase antibody type ELISA: an anti-hormone monoclonal antibody (Mab) is combined with a rabbit anti-mouse Ig antibody (RAMIG) that has been adsorbed on a solid phase carrier, and then added to a hormone standard or a sample to be tested, thereby The solidified Mab is combined with horseradish peroxidase (HRP) labeled hormone (enzyme-labeled hormone). By measuring the amount of bound enzyme hormone, the amount of hormone in the unknown sample can be converted. B. Solid phase antigen type ELISA: The "hormone-protein" complex (the protein should be different from the carrier protein in the immunogen) is coated on the solid phase carrier, and the hormone to be tested (sample or standard) and anti-IAA are added. The antibody (Pab) was cloned and subjected to a competition reaction. Then, HRP-labeled goat anti-rabbit Ig antibody (HRP-GARIG) was reacted with Pab bound to the solid phase, and the amount of the hormone in the unknown sample was converted by measuring the amount of the enzyme bound to the solid phase.
This chapter will explain the experimental steps and preparations of the plant hormone Elisa kit. Please click for details.
Our company needs spare materials, equipment and reagents when testing phytohormones:

(1) Materials: tissues or organs such as higher plants, fungi, algae.

(2) Instruments and equipment: 1. Enzyme-linked immunosorbent detector; 2. High-speed refrigerated centrifuge; 3. Thermostat; 4. Continuous injector; 5. Vortex; 6. 96-well microplate; 7. Centrifugation Tube; 8. mortar or homogenizer; 9. test tube.

(3) Reagents 1. Wash buffer: 0.01 mol/L pH 7.4 phosphate buffer (PBS), containing 0.05% Tween-0; 2. RAMIG solution, or "hormone-protein" complex; 3. 0.1% closed Protein (this protein should be different from the carrier protein in the immunogen); 4. Anti-hormone Mab, or Pab; 5. Hormone standard mother liquor, and reference series solution (diluted in double or quadruple series); 6. HRP Labeling hormone, or HRP-GARIG; 7. O-phenylenediamine (OPD) substrate solution: 5 mg of OPD was dissolved in 12.5 ml of 0.01 mol/L pH 5.0 PBS, and 30% H2O 212.5 μl was added before use.

According to our many years of experience in testing, we use solid phase antibody ELISA for the experimental manipulation of phytohormones.
First, the optimal working concentration of each reactant was selected by square matrix method; 100 μl of RAMIG solution was coated with polystyrene reaction plate micropores, 4 ° C wet box, 12 h;
Special attention must be made to discard the solution in the well, wash it 3 times with washing buffer, dry it; add 100μl anti-hormone Mab solution, 37 ° C for 70min; follow the above steps, it is not easy to have problems, thank you for Shanghai Jin Ma Bio's support, such as the phytohormone Elisa kit technical issues, can call us. Sincerely at your service.