Shanghai immunohistochemical technology experiment

Shanghai immunohistochemistry experiment

According to antigen antibody reactions and chemical color development principles, antigenic and primary anti-combination of tissue sections or cell specimens, then use an anti-marking biotin , Secondary antibodies such as fluorescein, the former, then combined with antibiotic (such as streptavidin, etc.) labeled horseradish peroxidase (HRP) or alkaline phosphatase (AKP), and then Color reaction or fluorescence showing the chemical composition in cells or tissues, clearly seeing the antigen antibody reaction product that occurs within the optical microscope or fluorescent microscope, thereby determining certain chemical components on the cell climb or tissue slice. Distribution and content. Shanghai immunohardization technology experiment

frozen slice 1, the cutting of frozen slices will be fixed with 4 ° C cold acetone 5 ~ 30min, PBS was washed 5 to 10 min 3 min × 3 2, and 0.3% Tritonx-100, 37 ° C was incubated for 20 to 40 min, PBS washed 5 to 10 min X3



[ 123] Paraffin Slices 1, put paraffin slices at 60 ° C for 1 h, then in conventional deprics of water
2, 0.3% Tritonx-100, 37 ° C for 20 ~ 40min, PBS Wash 5 ~ 10min x3 3, add 3% H2O2, 37 ° C for 5 to 30 min, PBS wash 5 to 10 min X3
4, dripping the osmolaized sealing liquid, 37 ° C Incubation 5 ~ 30min, do not wash
5, add appropriate proportion of dilution, 4 ° C incubation
6, take out the slice, place for 30 ~ 60 min, PBS 5 to 10 min X3
7, drop Galastinized secondary antibody, 37 ° C 20 to 40 min, PBS wash 5 to 10 min x3
8, drip hospordease labeled streptris, 37 ° C 20 ~ 40min, PBS wash 5 ~ 10 min x3 [ 123] 9, formulated DAB color color, 37 ° C color production 10-30min, thoroughly wash
10, Sukin reacted, water is washed back
11, alcohol dehydration, xylene transparent, neutral gum sealing [ 123] Customers provide
1 organization: fresh or fixed (10% neutral formaldehyde, 4% polyformaldehyde) tissue sample;
2 cells: fresh (greater than 10E6) or fixed (10% neutral formaldehyde, 4 % Polyfolform) cell sample;
3 Slices: paraffin slices, frozen slices and cell climbs, smear;
4 experimental materials: one-to-antif;
Biological Provide
1 Dye Slices;
2 electronic picture
3 Experiment Report (material, reagent, instrument, method, data analysis, experimental results);


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