笔记详情
标题
Immunohistochemistry
内容
Immunohistochemistry
After excising the tumor tissues and other organ tissue from mice, specimens were fixed in 10% formaldehyde for 48 hours and paraffin-embedded. Consecutive sections of 4 micron thick were cut and processed for hematoxylin and eosin staining; immunohistochemistry for pACC (cat. no. 3661, used at 1:100), p-mTOR (cat. No: 2976, used at 1:50), pAkt (Ser473, cat. No: 4060 used at 1:50), SIRT1 (cat no: 15404, used at 1:100) and Ki-67 (cat. No: ab15580, used at 1:100), ɣH2A.X (cat no:9718, used at 1:100). Antibodies to pACC, p-mTOR, pAkt, and ɣH2A.X were from Cell Signaling Technology (Denver, MA). Ki-67 was from Abcam (Cambridge, MA). p16 was from Proteintech (cat. No: 10883–1-AP, used at 1:200; Proteintech Group, Chicago, IL). SIRT1 was from Santa Cruz Biotech (Santa Cruz, CA). Solutions obtained from Dako Cytomation (Carpinteria, CA) were used for performing immunostaining as described before [13, 93]. The slides were examined under a light microscope, and representative pictures were taken from a minimum of 3 slides of each group [93]. The quantification of the stain intensity was performed by assigning a score of 0–1 for no or weak stain; 2 for moderate stain and 3 for strong stain. All slides were examined in a blinded manner by 2 individuals, including a pathologist (RA). 点击翻译
After excising the tumor tissues and other organ tissue from mice, specimens were fixed in 10% formaldehyde for 48 hours and paraffin-embedded. Consecutive sections of 4 micron thick were cut and processed for hematoxylin and eosin staining; immunohistochemistry for pACC (cat. no. 3661, used at 1:100), p-mTOR (cat. No: 2976, used at 1:50), pAkt (Ser473, cat. No: 4060 used at 1:50), SIRT1 (cat no: 15404, used at 1:100) and Ki-67 (cat. No: ab15580, used at 1:100), ɣH2A.X (cat no:9718, used at 1:100). Antibodies to pACC, p-mTOR, pAkt, and ɣH2A.X were from Cell Signaling Technology (Denver, MA). Ki-67 was from Abcam (Cambridge, MA). p16 was from Proteintech (cat. No: 10883–1-AP, used at 1:200; Proteintech Group, Chicago, IL). SIRT1 was from Santa Cruz Biotech (Santa Cruz, CA). Solutions obtained from Dako Cytomation (Carpinteria, CA) were used for performing immunostaining as described before [13, 93]. The slides were examined under a light microscope, and representative pictures were taken from a minimum of 3 slides of each group [93]. The quantification of the stain intensity was performed by assigning a score of 0–1 for no or weak stain; 2 for moderate stain and 3 for strong stain. All slides were examined in a blinded manner by 2 individuals, including a pathologist (RA). 点击翻译
来源
Zaid Al-Wahab. Oncotarget. 2015 Mar 12
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