Bcr-Abl fusion protein detection method
A bcr-abl, fusion protein technology, applied in measuring devices, instruments, material analysis by optical means, etc., can solve the problems of small number of chromosome karyotype analysis, easy to be affected by reaction conditions, high false positive rate, and save money Resources, easy separation and purification, high sensitivity
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Embodiment 1
[0023] The Bcr-Abl fusion protein detection method of the present embodiment, see figure 1 , 2 , including the following steps:
[0024] (1) Preparation of cell lysate: Take 8 mL of culture medium of K-562 cells (CML cells, Bcr-Abl fusion protein positive cells) with a cell concentration of 1.25×106 cells / mL, transfer it to a 10 mL centrifuge tube, and centrifuge at 500 g for 10 min to remove the culture medium. Then use 10 mL of cold PBS to wash by centrifugation at 500 g for 5 min, repeat twice, add 2 mL of prepared lysate (1 mL RIPA solution, 10 μL phosphatase inhibitor, 5 μL 100 mM PMSF solution, 1 μL protease inhibitor), and shake for 30 seconds Let stand on ice for 4 minutes, repeat 5 times, centrifuge to get the supernatant, store at -70°C in aliquots.
[0025] (2) Prepare gold-coated silver nanorods. The preparation method includes the following steps: first adopt CTAB-assisted gold seed growth method to prepare gold nanorods, centrifuge the synthesized gold nanorods...
Embodiment 2
[0036] The Bcr-Abl fusion protein detection method of the present embodiment comprises the following steps:
[0037] (1) Preparation of cell lysate: Take 8 mL of the culture medium of K-562 cells (CML cells, Bcr-Abl fusion protein positive cells) with a cell concentration of 1.25×106 cells / mL, transfer it to a 10 mL centrifuge tube, and centrifuge at 800 g for 10 min to remove the culture medium. Then use 10 mL of cold PBS to wash by centrifugation at 800 g for 5 min, repeat twice, add 2 mL of prepared lysate (1 mL RIPA solution, 10 μL phosphatase inhibitor, 5 μL 100 mM PMSF solution, 1 μL protease inhibitor), shake for 40 Seconds, let stand on ice for 4 minutes, repeat 8 times, centrifuge to get the supernatant, store at -70°C in aliquots. The lysis process of ccrf cells (acute lymphoid leukemia (ALL) cells, Bcr-Abl fusion protein negative cells) was the same as above.
[0038] (2) Prepare gold-coated silver nanorods. The preparation method includes the following steps: firs...
Embodiment 3
[0049] The Bcr-Abl fusion protein detection method of the present embodiment comprises the following steps:
[0050] (1) Preparation of cell lysate: Take 8 mL of culture medium of K-562 cells (CML cells, Bcr-Abl fusion protein positive cells) with a cell concentration of 1.25×106 cells / mL, transfer it to a 10 mL centrifuge tube, and centrifuge at 600 g for 10 min to remove the culture medium. Then use 10 mL of cold PBS to wash by centrifugation at 700 g for 5 min, repeat twice, add 2 mL of prepared lysate (1 mL RIPA solution, 10 μL phosphatase inhibitor, 5 μL 100 mM PMSF solution, 1 μL protease inhibitor), shake for 35 Seconds, let stand on ice for 4 minutes, repeat 7 times, centrifuge to get the supernatant, store at -70°C. The lysis process of ccrf cells (acute lymphoid leukemia (ALL) cells, Bcr-Abl fusion protein negative cells) was the same as above.
[0051] (2) Prepare gold-coated silver nanorods. The preparation method includes the following steps: first adopt CTAB-ass...
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