The preparation for the production of sections was processed as described previously (Xu et al., 2020 (link)). Then 3 μm sections at the site of femur metaphysis were cut coronally for Alcian Blue Hematoxylin/Orange G (ABH) staining. The indexes of histomorphometry including the area of lipid droplets and area of trabecular bone were detected with the use of OsteoMetrics software (Decatur, GA). IHC was performed using anti-alkaline phosphatase (ALP, ARIGO, ARG57422 1:300), anti-fatty acid-binding protein (FABP4, Abcam, ab92501, 1:200), anti-β-catenin (HuaBio, ER0805, 1:200). The quantitative analyses for positive staining area were performed by the software of image-pro plus6.0 (Media Cybernetics, Silver Spring, United States).
Arg57422
Manufactured by Arigo Biolaboratories
Sourced in United Kingdom
The ARG57422 is a high-precision laboratory centrifuge designed for a wide range of applications. It features a robust construction, a brushless motor, and a user-friendly control panel for easy operation. The centrifuge can accommodate various types of sample containers and offers adjustable speed and time settings to meet the specific needs of your laboratory procedures.
Automatically generated - may contain errors
3 protocols using arg57422
1
Histomorphometric Analysis of Femur Metaphysis
2
Quantifying Osteogenic Markers in Cell Cultures
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Food & Function 948 | Food Funct., 2023, 14, 946-960
Huabio, Hangzhou, Zhejiang, China), runt-related transcription factor-2 (Runx2; diluted 1 : 1000, ET1612-47; Huabio, Hangzhou, Zhejiang, China), alkaline phosphatase (ALP; diluted 1 : 5000, ARG57422; Arigo, Taiwan, China), peroxisome proliferator-activated receptor gamma (PPARγ; diluted 1 : 1000, ARG55241; Arigo, Taiwan, China), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; diluted 1 : 5000, ET1601-4; Huabio, Hangzhou, Zhejiang, China) in tris-buffered saline Tween-20. Then, the membranes were washed and incubated with the corresponding secondary antibody at room temperature without light for 2 h. Then, the expression of proteins was detected using an Odyssey® CLX Imaging System (LI-COR Biosciences, Nebraska, USA) following the manufacturer's protocol.
Huabio, Hangzhou, Zhejiang, China), runt-related transcription factor-2 (Runx2; diluted 1 : 1000, ET1612-47; Huabio, Hangzhou, Zhejiang, China), alkaline phosphatase (ALP; diluted 1 : 5000, ARG57422; Arigo, Taiwan, China), peroxisome proliferator-activated receptor gamma (PPARγ; diluted 1 : 1000, ARG55241; Arigo, Taiwan, China), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; diluted 1 : 5000, ET1601-4; Huabio, Hangzhou, Zhejiang, China) in tris-buffered saline Tween-20. Then, the membranes were washed and incubated with the corresponding secondary antibody at room temperature without light for 2 h. Then, the expression of proteins was detected using an Odyssey® CLX Imaging System (LI-COR Biosciences, Nebraska, USA) following the manufacturer's protocol.
3
Femur Morphology and Angiogenesis Analysis
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After Micro-CT scanning, femur samples were processed for para n sections at the thick of 3µm as previously described 17 . Alcian Blue Hematoxylin/Orange G (ABH) staining were performed on these sections for morphological analysis. The numbers of blood vessel and trabecular area (%) in the region of interest were measured using OsteoMetrics software (Decatur, GA, USA) by two researchers. The IHC assay were detected as follows: 1. sections were treated with 0.01 M citrate buffer (Solarbio, Beijing, CN) at 60°C for 4 h as antigen retrieval; 2. sections then were incubated in primary antibodies of CD31 (Diagbio, AGR52748, CN), VEGF (ARIGO, ARG10513, CN), β-catenin (abcam, ab32572, UK), alkaline phosphatase (ALP, ARIGO, ARG57422, CN) overnight at 4°C; 3. sections were incubated in secondary antibodies for 20 min and diaminobenzidine (DAB) solution for 1 min to detect positive staining; 4. sections were counterstained with hematoxylin. Image_J software was used to analyzed the quanti cation of positive staining.
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