Immunohistochemical Analysis of Spermatogenesis

Bouin's fixed 5-μm thick sections were processed for immunohistochemistry. Staining was carried out using a Lab Vision PT module (Thermo Fisher Scientific) and Autostainer 480S (medac) as described previously (Umer et al., 2021 (link)). Primary antibodies against HOOK1 (1:200; 10871-1-AP) (Zhou et al., 2009 (link)), LC3B (1:100; ab58610, Abcam), ARL3 (1:200; 10961-1-AP) (Qi et al., 2013 (link)), GOPC (1:200; 12163-1-AP, Thermo Fisher Scientific), COPA (1:200; BS-12464R, Thermo Fisher Scientific), PICK1 (1:200; PA5-76084, Thermo Fisher Scientific), SP56 (1:200; MA1-10866, Thermo Fisher Scientific), acrosin (1:200; BS-5151R, Thermo Fisher Scientific), AKT1 (1:100; 4060, Cell Signaling Technology), SQSTM1 (1:200; 5114, Cell Signaling Technology) and ARF3 (1:200; 10800-1-AP, Proteintech) were used.

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Umer N., Phadke S., Shakeri F., Arévalo L., Lohanadan K., Kirfel G., Sylvester M., Buness A, & Schorle H. (2022). PFN4 is required for manchette development and acrosome biogenesis during mouse spermiogenesis. Development (Cambridge, England), 149(16), dev200499.

Publication 2022

Autostainer 480S ab58610

Acrosin Akt1 Antibodies Copa Immunohistochemistry Vision

Corresponding Organization : University Hospital Bonn

Other organizations : University of Bonn, Institut für Medizinische Biometrie, Informatik und Epidemiologie

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Variable analysis

independent variables

Primary antibodies against HOOK1, LC3B, ARL3, GOPC, COPA, PICK1, SP56, acrosin, AKT1, SQSTM1, and ARF3

dependent variables

Immunohistochemistry staining

control variables

Bouin's fixed 5-μm thick sections
Lab Vision PT module (Thermo Fisher Scientific) and Autostainer 480S (medac) for staining procedure

controls

Positive controls not explicitly mentioned
Negative controls not explicitly mentioned

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