Plk3 prostatic cancer prognosis detection reagent and kit
A detection kit and a technology for prostate cancer, applied in the biological field, can solve the problems of unpredictable patient metastasis, unknown expression and biological role of Plk3 in prostate cancer, insufficient specificity, etc.
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[0030] The method of using this kit is as follows:
[0031] (1) Place the slices that have been dewaxed, hydrated, and antigen-restored in the incubation box, and circle the tissue with a water-blocking pen to prevent the reagents from dripping from spreading, and absorb the water on the slices with absorbent paper. , do not touch the tissue, add H 2 o 2 Cover the tissue with 50ul and incubate at room temperature for 30min to block the activity of endogenous peroxidase;
[0032] (2) Rinse with PBS for 3 minutes, repeat 3 times;
[0033] (3) Slightly absorb the PBS solution on the slice with absorbent paper, add 50ul non-immune sheep serum working solution dropwise, and incubate at room temperature for 15 minutes;
[0034] (4) Gently shake off the serum without rinsing, add dropwise 50ul of primary antibody (2μg / ml rabbit-derived anti-Plk3 polyclonal antibody) (Plk3 antibody:PBS=1:100), incubate at room temperature for 60 minutes or overnight at 4°C;
[0035] (5) Rinse wit...
experiment example
[0042] Experimental example: Selection experiment of markers indicative of proliferation and metastasis of prostate cancer
[0043] 1: The expression of Plk3 in prostate cancer and adjacent tissues
[0044] The expression of Plk3 in 68 cases of prostate cancer and 10 cases of normal prostate tissue was detected by immunohistochemical chip. Such as figure 1As shown in A-F, Plk3 is strongly positive in prostate cancer cell plasma and moderately positive in normal prostate tissue. Among the 68 prostate cancer tissues, 19 cases (27.9%) had low expression of Plk3, and 49 cases (72.1%) had high expression of Plk3. And the expression level of Plk3 in prostate cancer tissue was significantly higher than that in normal prostate tissue (IRS: PCa=4.98±0.95vs.Normal=4.30±0.82, P=0.034; T1-T2=4.55±0.90vs.T3-T4= 5.61±0.63, Pfigure 1 B).
[0045] Samples are now fixed in 10% neutral formalin and then embedded in paraffin. Paraffin sections were 4 μm thick, dewaxed in xylene, hydrated wi...
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