Immunofluorescence Staining of Newborn Mouse Femur

The prepared femur sections of newborn C57BL/6 mice were subjected to antigen retrieval with heat-induced antigen retrieval solution (ab973, abcam). The specific IF stain procedure was performed as outlined previously 33 (link). Primary antibodies, goat anti-GPR125 (1:200) (Santa Cruz) and mouse-anti-Ctsk (Santa Cruz, sc-48353) were applied to one section. After overnight incubation, secondary antibodies, TRITC donkey anti-Goat antibody (Thermo Scientific) and FITC donkey-anti-mouse antibody (Thermo Scientific) were used to show the specific expression location. Nuclei were visualized with DAPI (1ug/mL).

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Tang C.Y., Wang H., Zhang Y., Wang Z., Zhu G., McVicar A., Li Y.P, & Chen W. (2022). GPR125 positively regulates osteoclastogenesis potentially through AKT-NF-κB and MAPK signaling pathways. International Journal of Biological Sciences, 18(6), 2392-2405.

Publication 2022

ab973

Anti antibody Antibodies Antigen C57bl mice Ctsk Dapi Donkey Femur Fitc Goat Mouse Newborn Nuclei Stain Tritc

Corresponding Organization :

Other organizations : University of Alabama at Birmingham, Xidian University, Tulane University

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Variable analysis

independent variables

Type of primary antibody used (goat anti-GPR125, mouse anti-Ctsk)

dependent variables

Specific expression location of GPR125 and Ctsk proteins

control variables

Preparation of mouse femur sections
Antigen retrieval method (heat-induced antigen retrieval solution)
Incubation time for primary and secondary antibodies
Concentration of DAPI used to visualize nuclei

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

Annotations

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