Immunohistochemical Analysis of Liver Sections

Fresh-frozen liver sections were incubated with antibody to C1q (ab11861, Abcam, 1:50 dilution), and formalin-fixed, paraffin embedded liver sections were incubated as described (23 (link)) with antibodies to C3d (AF2655, R&D, 1:500 dilution), MAC (ab55811, Abcam, 1:800 dilution), C1q (ab11861, Abcam, 1:50 dilution), MPO (PA5-16672, Thermo Fisher, 1:400 dilution), 3-chlorotyrosine (HP5002, Hycult Biotech, 1:200 dilution), F4/80 (70076s, CST, 1:500 dilution), and VCAM-1 (MA5- 31965, Thermo Fisher, 1:400 dilution), followed by incubation with secondary antibody (HRP-polymer detection kit PV-9001, PV-9003 or PV-9004, Zsbio), according to the manufacturer’s instructions. Immunohistochemistry results were graded based on the staining intensity and percent positive cells as described (24 (link)).

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Guo Z., Chen J., Zeng Y., Wang Z., Yao M., Tomlinson S., Chen B., Yuan G, & He S. (2022). Complement Inhibition Alleviates Cholestatic Liver Injury Through Mediating Macrophage Infiltration and Function in Mice. Frontiers in Immunology, 12, 785287.

Publication 2021

ab11861 AF2655 ab55811 PA5-16672 HP5002 PV-9001 PV-9003 PV-9004

3 chlorotyrosine Antibodies Antibody Cells Dilution Formalin Frozen sections Immunohistochemistry Liver Paraffin Polymer Vcam 1

Corresponding Organization : Guangxi Medical University

Other organizations : Medical University of South Carolina

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Variable analysis

independent variables

Antibody used (C1q (ab11861, Abcam, 1:50 dilution), C3d (AF2655, R&D, 1:500 dilution), MAC (ab55811, Abcam, 1:800 dilution), C1q (ab11861, Abcam, 1:50 dilution), MPO (PA5-16672, Thermo Fisher, 1:400 dilution), 3-chlorotyrosine (HP5002, Hycult Biotech, 1:200 dilution), F4/80 (70076s, CST, 1:500 dilution), and VCAM-1 (MA5-31965, Thermo Fisher, 1:400 dilution))
Tissue preparation (fresh-frozen liver sections, formalin-fixed, paraffin embedded liver sections)

dependent variables

Immunohistochemistry results (graded based on the staining intensity and percent positive cells)

control variables

Incubation conditions (as described in the referenced publication (23))

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