Protein samples from the thoracic aorta were prepared in lysis buffer as described previously [11 (link), 41 (link)–47 (link)]. The proteins (15 μg) were separated by SDS–PAGE and transferred onto nitrocellulose membrane (Bio-Rad). The membrane was then incubated overnight (4 °C) with a primary antibody against Collagen I (1:1000, Sigma–Aldrich), elastin (1:200, Santa Cruz Biotechnology), TGFβ−1(1:100, Santa Cruz Biotechnology), scleraxis (1:100, Santa Cruz Biotechnology), MMP-2, MMP-9 (1:1000, EMD Millipore Corporation), Klotho (R&D Systems), and β-actin (1:10,000, Abcam). Goat anti-mouse or goat anti-rabbit conjugated with horseradish peroxidase (1:3000–1:5000, Santa Cruz Biotechnology) was used as a secondary antibody and incubated for 1 h at room temperature. Specific proteins were detected by chemiluminescent methods using Amersham™ ECL™ Western blotting detection reagents (GE Healthcare, UK). Protein abundance on Western blots was captured using Chem-Doc (Bio-Rad) and quantified by densitometry using the Image lab software (Bio-Rad, Hercules, CA).
Scleraxis
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Scleraxis is a high-quality lab equipment product designed for use in scientific research. It serves as a tool for monitoring and analyzing the expression of the scleraxis gene, which plays a crucial role in the development and differentiation of tendon and ligament tissues. The core function of Scleraxis is to provide researchers with accurate and reliable data on the expression patterns of this important regulatory gene.
Automatically generated - may contain errors
Lab products found in correlation
Elastin, by Santa Cruz Biotechnology (1 mentions)
Klotho, by R&D Systems (1 mentions)
Chemdoc, by Bio-Rad (1 mentions)
Tgf β1, by Santa Cruz Biotechnology (1 mentions)
Amersham ecl western blotting detection reagent, by GE Healthcare (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
β actin, by Abcam (1 mentions)
Mmp 2, by Merck Group (1 mentions)
Mmp 9, by Merck Group (1 mentions)
Apotome fluorescence microscope, by Zeiss (1 mentions)
Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions)
Collagen type 1, by Merck Group (1 mentions)
2 protocols using scleraxis
1
Quantitative Analysis of Aortic Extracellular Matrix Proteins
2
Tenocyte Marker Expression Analysis
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Expression of the tenocyte markers scleraxis (Santa Cruz Biotechnology, Dallas, Texas, USA), collagen type I (Merck Millipore, Milano, Italy) and the proliferative marker PCNA (Santa Cruz Biotechnology, Dallas, Texas, USA) was assessed by immunofluorescence. Primary monoclonal antibodies were diluted at 1 : 200 in PBS-1% BSA and incubated with the sections for 2 h at room temperature. The secondary DyLight 488 antibody (KPL, Kirkegaard & Perry Laboratories, Maryland, USA), diluted at 1 : 100, was incubated for 1 h at room temperature. Stained sections were visualized with an Apotome fluorescence microscope (Zeiss). We collected digital images using a ×20 dry lens within 0–5 days after labeling.
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