An antibody chip kit for simultaneous quantitative detection of multiple receptors
An antibody chip and quantitative detection technology, applied in the field of biomedicine, can solve the problems of cumbersome operation, low throughput, low sensitivity, etc., and achieve the effect of overcoming cumbersome operation and less specimen consumption
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Embodiment 1
[0024] Example 1: Screening of the best antibody chip carrier.
[0025] Conventional antibody chips mostly use nitrocellulose membrane as the carrier. Because the nitrocellulose membrane has a multi-layer structure, it is difficult to clean the chip, so that the background of the chip is high and the result is large. At the same time, the nitrocellulose membrane is fragile and not easy to operate on a large scale, and the use of large-scale clinical samples is not widespread. Different manufacturers use different methods to fix the nitrocellulose membrane on the surface of the glass slide. Among them is Whatman's SS glass slide, which fixes 16 cells containing nitrocellulose membrane on a glass slide. In addition, Gentel has used processing The nitrocellulose material is coated on the surface of the glass slide to produce the PATH glass slide. In addition, in order to fix the antibody on the surface of the glass slide, we screened the carrier activated by different methods. Spo...
Embodiment 2
[0028] Example 2: Preparation of an antibody chip kit for the simultaneous quantitative detection of multiple receptors.
[0029] In order to detect whether there are corresponding receptors in the sample, prepare slides fixed with specific antibodies against the following proteins: tumor necrosis factor receptor 4-1BB, leukocyte activation adhesion factor, stimulating molecule B7-1, B cell maturation factor , Myeloid cells are specifically rich in leucine glycoprotein, tumor necrosis factor receptor CD30, tumor necrosis factor ligand-5, carcinoembryonic antigen-related cell adhesion molecule 1, death receptor 6, Axl / Ufo growth factor receptor Family tyrosine kinase, cell membrane glycoprotein, receptor type tyrosine kinase ErbB3, E-selectin, tumor necrosis factor receptor superfamily member 6, human FMS-like tyrosine kinase 3 ligand, glucocorticoid-induced Tumor necrosis factor receptor family related proteins, herpes virus invasion mediator, intercellular adhesion molecule 3, i...
Embodiment 3
[0047] Example 3: Experiments for quantitative detection of receptors using the kit of the present invention.
[0048] 1. Complete drying of the slide chip
[0049] Take the slide chip out of the box, and after equilibrating at room temperature for 20-30 minutes, open the packaging bag, remove the sealing strip, and then place the chip in a vacuum desiccator or dry at room temperature for 1-2 hours.
[0050] 2. Press image 3 Perform a serial dilution of the cytokine standard gradient as shown in
[0051] 2.1. Add 500μl of sample diluent to the small tube of cytokine standard mixture, and re-dissolve the standard. Before opening the small tube, centrifuge it quickly, gently tap up and down to dissolve the powder, and mark this small tube as Std1.
[0052] 2.2. Mark 6 clean centrifuge tubes as Std2, Std3 to Std7, and add 200μl of sample diluent to each tube.
[0053] 2.3. Extract 100μl of Std1 and add it to Std2 and mix gently, then extract 100μl from Std2 and add it to Std3, so as to g...
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