Immunofluorescence kit for detecting human prostate cancer antigens PSMA and AR-V7 and application
A technology for prostate cancer and immunofluorescence detection, applied in biological testing, material inspection products, etc., can solve the problems of repeated sampling difficulties, a single tissue sample is not enough to reflect the tumor burden, and cannot be dynamically monitored
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Embodiment 1
[0080] Materials: smears of negatively enriched blood samples, and 22RV1 and LNCaP cell smears for control cells.
[0081] Experimental steps:
[0082] 1. Draw 3.5ml of peripheral blood into an ACD (sodium citrate) anticoagulant tube. use The human peripheral blood leukocyte depletion kit negatively enriches tumor cells and fixes them on glass slides;
[0083] 2. Wash slides with CYP1 for 3 minutes x 3 times, 100-150 μL each time, to ensure that the entire sample area is covered;
[0084] 3. Absorb the excess liquid on the slide, add CYPP for 5 minutes, wash the slides with CYP1 as above for 3 minutes × 1 time; absorb excess liquid, add 200 μl of ice acetone:methanol (7:3) for 5 minutes, and wash the slides with CYP1 for 3 minutes × 3 times , to absorb excess water;
[0085] 4. Add 100-150 μl of blocking solution to block at room temperature for 25-30 minutes. Absorb excess blocking solution, add 100 μl of diluted PSMA antibody, AR-V7 antibody and CD45 antibody, and incu...
Embodiment 2
[0095] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by membrane filtration and then detected for protein. Experimental steps:
[0096] 1. Take an appropriate amount of peripheral blood and put it into a blood collection tube containing anticoagulant, and shake it slightly to mix.
[0097] 2. Add the suspension to the membrane filtration separation tumor cell technology device, and slowly pass through the filter and the filter membrane.
[0098] 3. After the filtration is completed, continue to add 50ml of 0.01M PBS to the membrane filtration device, wash the cell suspension attached around the tube wall into the membrane filtration device, and let it pass through the filter and membrane;
[0099] 4. Fix the cells on the filter membrane;
[0100] 5. Perform the same operation as in Example 1 to detect the protein.
Embodiment 3
[0102] Materials: 1 tube of appropriate amount of anticoagulated blood, which is enriched by microfluidic method and then detected for protein
[0103] Experimental steps:
[0104] 1. The appropriate amount of blood drawn is enriched using microfluidic chips of various principles.
[0105] 2. After enrichment, the samples were subjected to protein immunofluorescence detection.
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