Proteins from lysed cells or immunoprecipitates were denatured and loaded on sodium dodecyl sulfate polyacrylamide gels and then transferred to polyvinylidene difluoride membranes (Bio-Rad). After blocking with 5% (w/v) BSA or milk the membrane was incubated with the corresponding primary and secondary antibodies (Bio-Rad). The specific bands were analyzed using Thyphoon or ChemiDoc Imaging Systems (Bio-Rad).
Primary and secondary antibodies
Manufactured by Bio-Rad
Sourced in United States
Primary and secondary antibodies are essential tools used in various laboratory techniques, such as Western blotting, immunohistochemistry, and flow cytometry. Primary antibodies are specific to target proteins or antigens, while secondary antibodies are designed to bind to the primary antibodies, enabling detection and visualization of the target. These antibodies are available in a range of host species, isotypes, and conjugation options to accommodate specific experimental requirements.
Automatically generated - may contain errors
2 protocols using primary and secondary antibodies
1
Western Blot Protein Analysis
2
Bone Histology Analysis Protocol
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Intercepted bone mass was thawed, xed in 10% neutral formalin for 48 hours, and embedded in para n after decalci cation in 10% EDTA solution (Zhongshan Jinqiao, Beijing, China). The tissues were then cut into 5-µm thick sections and treated with 3% hydrogen peroxide for 10 minutes. Afterward, the sections were rinsed with phosphate-buffered saline, incubated with primary and secondary antibodies (Bio-Rad, Hercules, USA) sequentially, and exposed to DAB (TaKaRa, Dalian, China). The sections were counterstained with hematoxylin solution (TaKaRa, Dalian, China). Ten visual elds were randomly selected and observed under a high magni cation microscope (400×) (Leica DM500, Solms,germany), and the number of positively stained cells was calculated.
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