Protein chip used for detecting functional injury of central nervous system and manufacturing method therefor
A technology of central nervous system and protein chip, applied in the biological field, can solve the problems of large amount of data and high cost, and achieve the effect of simple research work, accelerated speed and process.
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Embodiment 1
[0039] Embodiment 1 (protein antibody chip):
[0040] a. Nerve injury-specific protein factor antibody:
[0041] Monoclonal antibodies to homologous protein factors (markers of nerve injury) of 381 specific genes closely related to central nervous system injury listed in Table 1 were used as central nervous system injury-specific protein factor antibodies.
[0042] b. Extraction of proteins in patient blood, cerebrospinal fluid or damaged tissue samples:
[0043] Use RIPA solution (1% Igepal CA-630, 0.5% Sodium deoxycholate, 0.1% SDS and PBS, add 10mMbeta-mercaptoethanol, 10ug / ml PMSF, 5ug / ml aprotinin for the extracted patient’s blood, cerebrospinal fluid or damaged tissue samples) , 0.1mg / ml benzamidine, 1ug / ml pepstatin A, 1ug / ml leupeptin, 100mM sodium orthovanadate and 15ul / ml Triton X-100) extract protein, homogenate and centrifuge and store at -70°C.
[0044] c. Fabrication of protein antibody chips and fixation of nerve injury-specific protein factor antibodies:
[...
Embodiment 2
[0052] Embodiment 2 (protein antigen chip)
[0053] a. Central nervous system injury-specific protein antigen:
[0054] The homologous protein-specific proteins (which can be ordered from other companies) of 381 genes closely related to central nervous system injury listed in Table 1 were used as central nervous system injury-specific protein antigens.
[0055] b. Patient serum or cerebrospinal fluid samples:
[0056] The patient's blood or cerebrospinal fluid samples were taken, centrifuged to obtain serum or cerebrospinal fluid supernatant, and stored at -70°C for later use.
[0057] c. Fabrication of protein antigen chips and fixation of nerve injury-specific protein antigens:
[0058] Using a high-speed automatic microarray spotting machine (CM-1000), 381 kinds of specific protein antigens closely related to central nervous system injury listed in Table 1, positive control protein antigens (Caspases, Tumor necrosis factor, etc.), negative control protein antigens ( Huma...
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