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Embodiment 1
[0028] A preparation method for rat CD4 antibody-coated magnetic beads, said method comprising the following steps:
[0029] Step 1: Wash the epoxy resin magnetic beads with an appropriate amount of 75% acrolein buffer reagent and vortex. After washing, put it on the magnet for 1min, and then remove the supernatant. In this process, as long as the amount of acrolein buffer can be used to wash the epoxy resin magnetic beads, it does not matter if it is too much.
[0030] Step 2: After adding acrolein buffer and PBS buffer to the epoxy resin magnetic beads pretreated in step 1, centrifuge at room temperature at 37°C for 16 hours; the proportion of each component is as follows: 5 mg epoxy resin magnetic beads Beads, 100ul rabbit anti-mouse antibody, 150ul acrolein buffer, 250ul PBS buffer.
[0031] Step 3: The epoxy resin magnetic beads treated in step 2 were precipitated by magnet for 1 min, and then washed by HB reagent; 5mg of epoxy resin magnetic beads were washed with 200u...
Embodiment 2
[0037] A kit, comprising the rat CD4 antibody-coated magnetic beads prepared by the method for preparing rat CD4 antibody-coated magnetic beads in Example 1.
Embodiment 3
[0039] ATP detection was carried out on the rat CD4 antibody-coated magnetic beads prepared by the invention, and the specific experimental process was as follows:
[0040] 1. First part of the test: cell stimulation.
[0041] Take out the relevant components in the kit from the refrigerator in advance and equilibrate to room temperature.
[0042] 1.1 Carefully mix the quality control control and rat CD4 whole blood samples by inversion, and repeat several times to ensure that the blood cells are evenly distributed. :
[0044] 1.3 After dilution, use one 8-well culture plate strip for each sample and assemble the culture plate according to the worksheet.
[0045] 1.4 Add 25ul of sample dilution to the first 4 wells of each strip as non-stimulation wells.
[0046] 1.5 Add 25ul of the sample dilution to the remaining 4 wells of each...
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Abstract
The invention provides a rat CD4 antibody coated magnetic beads, a preparation method and an application therefor and a kit containing magnetic beads. The method for preparing the magnetic beads comprises the following steps of: washing the magnetic beads multiple times through different reagents, and removing the supernatant. The rat CD4 antibody coated magnetic beads can be prepared according tothe preparation method. The prepared rat CD4 antibody coated magnetic beads can be used in the kit, and used for detecting immunological rejection after rat kidneytransplantation. Compared with theprior art, the invention has the advantages that the rat CD4 antibody coated magnetic beads prepared by the method for preparing the rat CD4 antibody coated magnetic beads is combined with an immune cell function measurement kit ImmuKnow TW-Cylex , and can be used for detecting the immunological rejection after the rat kidneytransplantation.
Description
technical field [0001] The invention relates to a rat CD4 antibody-coated magnetic bead, a preparation method and application thereof, and a kit containing the magnetic bead. Background technique [0002] Currently on the market, there is no kit that can be used to detect immune rejection after kidneytransplantation in rats. Contents of the invention [0003] The object of the present invention is to provide a rat CD4 antibody-coated magnetic bead, its preparation method and application, and a kit containing the magnetic bead. [0004] The purpose of the present invention is achieved through the following technical solutions: [0005] A preparation method for rat CD4 antibody-coated magnetic beads, step 1: wash the magnetic beads with C1 reagent, then place them on a magnet for adsorption and remove the supernatant; [0006] Step 2: After adding antibody, C1 reagent, and C2 reagent to the magnetic beads pretreated in step 1, centrifuge at room temperature for 16-24 hour...
Claims
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