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Localization of NANOG protein expression was performed on mouse testis sections using immunohistochemistry and expression was detected in the nucleus of pachytene spermatocytes and round spermatids (Fig. 2A C).
Similarly, we noted PCNA staining of spermatogonia and leptotene spermatocytes, but not pachytene spermatocytes in adult mouse testis sections (Fig. S1A), which is consistent with a previous report [54].
Immunohistochemical staining was performed on 5- μm mouse testis sections, previously fixed in 4% formaldehyde and embedded in paraffin.
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(B ) Immunofluorescence on sections of mouse testis tubules.
(D ) Immunohistochemistry stainings (IHC) on sections of mouse testis tubules.
(A ) Immunohistochemistry (IHC) stainings on sections of mouse testis tubules for different acetylation sites on H3.
Paraffin sections of mouse testis (Novagen) were deparaffinized using xylene, rehydrated through a graded series of 100%, 95 %, 80, and 70% ethandl, and incubated in 3% hydrogen peroxide in methanol to quench endogenous peroxidase activity.
In order to define the cells in which CCDC6 is expressed in the normal testis, immunohistochemical analysis for this protein was performed on serial sections of the mouse testis.
To achieve this aim, analysis for CCDC6 expression has been evaluated on serial sections of the mouse testis by immunohistochemistry and on separate populations of murine testicular cells by western blot.
To confirm and further examine the developmental expression and localization of the five proteins in spermatogenic cells or mature sperm, we performed indirect immunofluorescence analysis in paraffin sections of adult mouse testis with the antibodies.
Paraffin-embedded testis sections (6 μm) of normal adult mouse (C57BL/6 mouse, male, 8 weeks) were obtained from Genostaff Co., Ltd.
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