The Opal 7 multiplexed assay (PerkinElmer, MA, USA) was used to generate multiple immunostainings. The best concentration of antibodies was determined before multiple immunostainings, according to the instructions. The primary antibodies used were as follows: anti-CD45 (1:200, ab10558, Abcam), anti-α-SMA (1:2000, ab12964, Abcam), anti-VIM (1:150, ab8978, Abcam), anti-CD31 (1:50,ab9498,Abcam), anti-NINJ1 (1:200, GTX31596, GeneTex), anti-TPPP3 (1:200, GTX33554, GeneTex), anti-CD3 (1:200, 17,617–1-P,Thermo), anti-MMP9 (1:1000, ab74003,Abcam), anti-CD11b (1:3000, ab133357, Abcam), anti-CD31 (1:50, ab28364,Abcam), anti-GSDMD (1:800, 36,425, Cell signaling), anti-HMGB1(1:500, ab79823, Abcam). The stained slides were analyzed by Vectra Polaris Quantitative Pathology Imaging Systems (PerkinElmer).
Ab74003
Manufactured by Abcam
Sourced in United States
Ab74003 is a recombinant anti-MAP1LC3B antibody. It is designed for the detection of MAP1LC3B protein.
Automatically generated - may contain errors
2 protocols using ab74003
1
Multiplex Immunostaining for Cellular Profiling
2
Immunohistochemical Localization of OSR1
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Immunohistochemistry was performed by Histostain-Plus Kits (Broad Spectrum) and an Invitrogen 2nd Generation LAB-SA Detection System (Invitrogen; Life Technologies). Slides were incubated with a primary antibody to OSR1 (ab74003; Abcam, Cambridge, MA, USA) followed by a biotinylated secondary antibody and enzyme conjugate, stained with 3,3'-diaminobenzidine, and finally counterstained with haematoxylin. The extent of OSR1 staining was scored by assigning the percentage of positive tumour cells (0, none; 1, < 20% of positive staining cells; 2, 20–50% of positive staining cells; 3, > 50% of positive staining cells).
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