Biglycan

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Biglycan is an extracellular matrix proteoglycan produced by Santa Cruz Biotechnology. It plays a role in the structural integrity and function of connective tissues.

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Lab products found in correlation

Catalase, by Santa Cruz Biotechnology (1 mentions) Mini protean tgx stain free, by Bio-Rad (1 mentions) Hybond c extra nitrocellulose membranes, by Bio-Rad (1 mentions) Superoxide dismutase 1 sod 1, by Cell Signaling Technology (1 mentions) Enhanced chemi luminescence (ecl), by Cytiva (1 mentions) Image lab software, by Bio-Rad (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Type 1 collagen, by Abcam (1 mentions) α sma, by Merck Group (1 mentions) Dm2000, by Leica (1 mentions) E cadherin, by Agilent Technologies (1 mentions) Enhanced chemiluminescence western blot detection system, by Cytiva (1 mentions) Snail, by Biorbyt (1 mentions) Peroxidase conjugated igg antibodies, by Medical & Biological Laboratories (1 mentions) β actin, by Abcam (1 mentions) Odyssey imaging system, by LI COR (1 mentions) 4 hne, by Abcam (1 mentions) Pvdf membrane, by LI COR (1 mentions) Nanodrop microvolume spectrophotometer, by Thermo Fisher Scientific (1 mentions) Hmgb1, by Abcam (1 mentions)

4 protocols using biglycan

Protein Expression Analysis in Aortic Valve

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Aliquots of 5-7.5 µg of total proteins were prepared and electrophoresed from AV (N = 71) and VICs extracts on reducing SDS polyacrylamide gels (4–15% polyacrylamide, Mini-PROTEANTGX Stain-Free, BioRad) and transferred to Hybond-C Extra nitrocellulose membranes (BioRad). Primary antibodies used were superoxide dismutase 1 (SOD-1, Cell Signaling), fumarase, catalase and biglycan (all purchased from Santa Cruz Biotechnology). Secondary antibodies for mouse and rabbit were purchased from GE Healthcare. Stain free and β-actin were used as loading controls for blot normalization. Positive blots were detected with a chemiluminescence method (ECL, Amersham Biosciences) and images acquired with Chemidoc MP Imaging system (Bio-Rad). Semiquantitative analyses were performed by band densitometry using Image Lab software (Bio-Rad) and normalized data was expressed as arbitrary units (A.U.). All western blots were performed at least in triplicate for each experimental condition. Representative blots for markers assessed by western blot are shown in online supplemental material Figure S1.

Martín-Núñez E., Goñi-Olóriz M., Matilla L., Garaikoetxea M., Mourino-Alvarez L., Navarro A., Fernández-Celis A., Tamayo I., Gainza A., Álvarez V., Sádaba R., Barderas M.G., Jover E, & López-Andrés N. (2023). Influence of diabetes mellitus on the pathological profile of aortic stenosis: a sex-based approach. Cardiovascular Diabetology, 22, 280.

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Histological Analysis of Aortic Valve Cusps

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In a representative subset of aortic valve cusp specimen, histological analyses were performed. Aortic valves of patients without (n = 9) or with LVAD (n = 11) were snap‐frozen in cryo compound, and 5 μm sections were prepared. Sections were stained according to standard protocols for haematoxylin/eosin, Movat's pentachrome and von Kossa staining as previously described.¹⁴ Immunohistochemical staining for detection of α‐SMA (Sigma) and vimentin as well as staining for CD3, CD86, elastin, collagen type 1 (Abcam), and biglycan (Santa Cruz) were performed as previously described.¹⁵ Analyses and documentation were performed in a blinded fashion using pseudonyms for sample labelling with a Leica DM2000 and Leica LAS software Version 3.8.0.

Barth M., Mrozek L., Niazy N., Selig J.I., Boeken U., Sugimura Y., Kalampokas N., Horn P., Westenfeld R., Kröpil P., Aubin H., Lichtenberg A, & Akhyari P. (2021). Degenerative changes of the aortic valve during left ventricular assist device support. ESC Heart Failure, 9(1), 270-282.

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Immunoblot Analysis of Cellular Proteins

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Whole-cell lysates were prepared as previously described [59 (link)]. Lysates (20 μg) were subjected to immunoblot analysis using SDS-PAGE (12.5%), followed by electrotransfer onto nitrocellulose filters. The filters were incubated with primary antibodies, followed by peroxidase-conjugated IgG antibodies (Medical and Biological Laboratories). Anti-tubulin antibody was used to assess the protein levels loaded per lane (Oncogene Research Products, Cambridge, MA, USA). The immune complex was visualized using an Enhanced Chemiluminescence Western-blot detection system (Amersham, Aylesbury, UK). Antibodies for biglycan (Santa Cruz), phosphorylated AKT (phosphoSer473, Proteintech Group Inc. Rosemont, IL, USA), E-cadherin (DAKO), CLDN4 (clone 4D3) [57 (link)], Snail (Biorbyt, St Louis, MO, USA), and CD44 (Abcam) were used as primary antibodies. β-actin, detected by antibody (Abcam), was used as the loading control.

Fujiwara-Tani R., Sasaki T., Fujii K., Luo Y., Mori T., Kishi S., Mori S., Matsushima-Otsuka S., Nishiguchi Y., Goto K., Kawahara I., Kondoh M., Sho M, & Kuniyasu H. (2020). Diabetes mellitus is associated with liver metastasis of colorectal cancer through production of biglycan-rich cancer stroma. Oncotarget, 11(31), 2982-2994.

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Quantifying Protein and Inflammatory Markers in Necrotic Bone Fluid

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BCA protein assay (Pierce™ BCA Protein Assay Kit, Thermo Fisher Scientific) was used to measure the protein concentration of all NBF samples described above and the control BF (bone fluid) from normal bone. The DNA concentration of NBF was measured by NanoDrop Microvolume Spectrophotometers (Thermo Fisher Scientific). ELISA (Thermo Fisher Scientific) was used to measure IL1β, IL6, and TNFα protein levels in the control BF and NBF collected at different time points. Western blot analysis was used to compare the amount of HMGB1 (Abcam), Biglycan (Santa Cruz Biotechnology), 4-HNE (Abcam), RANKL (Abcam), and IL6 (Abcam) present in different NBF samples. In brief, SDS-PAGE was performed to separate proteins in obtained NBF samples using 12% polyacrylamide gel with equal amount of protein loaded in each well (40 micro gram). After transferring to PVDF membrane (LI-COR Biosciences), each individual component was detected by the corresponding primary antibody. Odyssey Imaging System (LI-COR Biosciences) was used to detect the signal produced by IRDye secondary antibody against primary antibody.

Deng Z., Ren Y., Park M.S, & Kim H.K. (2021). Damage Associated Molecular Patterns in Necrotic Femoral Head Inhibit Osteogenesis and Promote Fibrogenesis of Mesenchymal Stem Cells. Bone, 154, 116215.

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