Chondroitinase abc chabc

Manufactured by Merck Group

Sourced in United States

Chondroitinase ABC (ChABC) is a bacterial enzyme that cleaves chondroitin sulfate, a component of the extracellular matrix. It is commonly used in laboratory research applications.

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Lab products found in correlation

Anti v5 fitc antibody, by Thermo Fisher Scientific (1 mentions) C1 confocal microscope, by Nikon (1 mentions) Anti rptpβ, by BD (1 mentions) Anti olig2, by Merck Group (1 mentions) Anti sox2, by Abcam (1 mentions) Anti gapdh, by Abcam (1 mentions) Mouse anti paxillin antibody, by BD (1 mentions) Anti mbp, by Santa Cruz Biotechnology (1 mentions) Anti sall2, by Proteintech (1 mentions) Anti py118 paxillin, by Cell Signaling Technology (1 mentions) Anti ng2 proteoglycan, by Merck Group (1 mentions) Heparinase 3, by New England Biolabs (1 mentions) Heparinase 2, by New England Biolabs (1 mentions) Heparinase 1, by New England Biolabs (1 mentions) Hyaluronic acid, by Merck Group (1 mentions) Heparin, by Merck Group (1 mentions) Heparinase 3, by Merck Group (1 mentions) Sodium acetate, by Merck Group (1 mentions) Laemmli sample buffer, by Bio-Rad (1 mentions) β mercaptoethanol, by Merck Group (1 mentions)

Close spelling variants of this product

Chondroitinase ABC (149 citations) ChABC (40 citations) Chondroitinase (10 citations)

9 protocols using chondroitinase abc chabc

Immunofluorescence Staining of BL Tissues

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For immunofluorescence staining, confirmed BL tissues from the SEER Residual Tissue Repository from the United States was used. FFPE tissue was deparafinized and re-hydrated with a series of xylene and ethanol solutions. After blocking, the tissue was incubated with 50 nM rVAR2, followed by incubation with anti-V5-FITC antibody (Invitrogen). The specificity of rVAR2 binding was tested by outcompeting protein binding with 400 μg/ml CSA (Sigma). An additional specificity control was performed by treating the tissue with 0.5 U/ml Chondroitinase ABC (ChABC, Sigma) for 1 hour at 37 °C, before blocking. Samples were counterstained with DAPI.
Imaging was performed on a Nikon C1 confocal microscope with a 60× oil objective. Each image is a composite of the following channels: blue (nuclei), green (ofCS), red (auto-fluorescence of the tissue).

Agerbæk M.Ø., Pereira M.A., Clausen T.M., Pehrson C., Oo H.Z., Spliid C., Rich J.R., Fung V., Nkrumah F., Neequaye J., Biggar R.J., Reynolds S.J., Tosato G., Pullarkat S.T., Ayers L.W., Theander T.G., Daugaard M., Bhatia K., Nielsen M.A., Mbulaiteye S.M, & Salanti A. (2017). Burkitt lymphoma express oncofetal Chondroitin Sulfate without being a reservoir for placental malaria sequestration. International journal of cancer, 140(7), 1597-1608.

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Purification of Recombinant Protein Tyrosine Phosphatases

Cited 1 time

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Recombinant proteins corresponding to the entire intracellular regions (ICRs) of PTPRZ1, PTPRA, and PTPRM were expressed using a baculovirus-silkworm expression system, and purified as described^{6 (link)}. The ICRs of PTPRG, PTPRS, and PTPRB and the catalytic domains of PTPN1 and PTPN6 were expressed as glutathione-S-transferase (GST) fusion proteins from each pGEX plasmid in Escherichia coli strain BL21 (ref. 6 (link)). GST fusion proteins were purified by glutathione affinity chromatography as described^{43 (link)}. Chondroitinase ABC (chABC) was purchased from Sigma-Aldrich (catalog #C3667). Anti-PTPRZ-S, rabbit polyclonal antibodies against the extracellular region of PTPRZ was described previously (ref. 44 (link)). The following are the specificities and sources of the commercially available antibodies used in the present study: Anti-RPTPβ (a monoclonal antibody against the intracellular domain of PTPRZ1 receptors, #610179, BD Biosciences), anti-SOX2 (#ab97959, Abcam), anti-POU3F2 (#12137, Cell Signaling), anti-OLIG2 (#AB9610, Millipore), anti-SALL2 (#12679–1-AP, Proteintech Group), anti-GAPDH (#ab9482, Abcam) anti-phosphotyrosine (PY20; #ab16389, Abcam), and anti-pY118-paxillin (#2541, Cell Signaling), mouse anti-paxillin antibody (#610569, BD Bioscience), anti-MBP (#sc-13914, Santa Cruz Biotechnology), and anti-NG2 proteoglycan (#AB5320, Millipore).

Fujikawa A., Sugawara H., Tanaka T., Matsumoto M., Kuboyama K., Suzuki R., Tanga N., Ogata A., Masumura M, & Noda M. (2017). Targeting PTPRZ inhibits stem cell-like properties and tumorigenicity in glioblastoma cells. Scientific Reports, 7, 5609.

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Proteoglycan Isolation and Enzymatic Characterization

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Proteoglycans were isolated from the CM by precipitation with cationic detergent cetylpyridinium chloride, and the precipitates were dissolved in isopropanol. The polyanionic macromolecules were precipitated by adding sodium acetate-saturated ethanol and washed in ethanol. The extract was dissolved in ammonium acetate and then treated with chondroitinase AC (ChAC, Sigma-Aldrich), chondroitinase B (ChB, Sigma-Aldrich), chondroitinase ABC (ChABC, Sigma-Aldrich), heparinase I (NEB, Ipswich, MA, UK), heparinase II (NEB), or heparinase III (NEB) for further analysis.

Zhu Y., Lam A.K., Shum D.K., Cui D., Zhang J., Yan D.D., Li B., Xu W.W., Lee N.P., Chan K.T., Law S., Tsao S.W, & Cheung A.L. (2021). Significance of serglycin and its binding partners in autocrine promotion of metastasis in esophageal cancer. Theranostics, 11(6), 2722-2741.

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Chondroitinase ABC Treatment of U373 Astrocytes

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U373 astrocytes were digested with chondroitinase ABC (ChABC) (Sigma‐Aldrich, USA) to remove the chondroitin/dermatan sulfate structures. A confluent monolayer of U373 astrocytes was treated with a Tris HCL (200 mM) / sodium acetate (200 mM) buffer containing 10 U/ml chABC for 4 hr at 37°C and 5% CO².

Kamermans A., Planting K.E., Jalink K., van Horssen J, & de Vries H.E. (2018). Reactive astrocytes in multiple sclerosis impair neuronal outgrowth through TRPM7‐mediated chondroitin sulfate proteoglycan production. Glia, 67(1), 68-77.

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Modulation of Aβ42 Oligomer Toxicity

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The cultures were treated with different molecules in the presence of Aβ42 oligomers: (1) 100 units/mL heparin (Sigma, H3149), which compete with heparan sulfate proteoglycans for Aβ42 binding to reduce Aβ42 uptake; (2) 0.05 units/mL heparinase III (Sigma, H8891), which degrades heparan sulfate proteoglycans, (3) 0.2 mg/mL hyaluronic acid (Sigma), a type of ECM in brain perineuronal net; (4) 0.05 U/mL chondroitinase ABC (Chabc, Sigma), which degrades chondroitin sulfate proteogly-cans, for 72−96 h, respectively. The culture without treatment was used as the control. The treated samples were evaluated by Live/Dead assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, immunocytochemistry, etc. The culture supernatants were collected for lactate dehydrogenase (LDH) activity assay.

Bejoy J., Song L., Wang Z., Sang Q.X., Zhou Y, & Li Y. (2018). Neuroprotective Activities of Heparin, Heparinase III, and Hyaluronic Acid on the Aβ42-Treated Forebrain Spheroids Derived from Human Stem Cells. ACS biomaterials science & engineering, 4(8), 2922-2933.

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Chondroitinase Treatment of Amniotic Fluid Samples

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Immediately after harvesting, AF samples collected from fetuses at E14, E16, E18, and E21 were centrifuged at 4000 rpm for 10 min to remove cells and debris, then stored at −80°C until further analysis. Prior to Western blotting, equal volumes of AF from 3 randomly selected MMC or normal AF samples were combined and digested with 0.03 international units (IU) of chondroitinase ABC (ChABC; Sigma Aldrich, Saint Louis, MO, USA) in 0.1 M Tris-HCl buffer (pH = 8.0) containing 0.03 M sodium acetate and protease inhibitor cocktail (1:100; Sigma Aldrich, Saint Louis, MO, USA) for 3 h at 37 °C with gentle shaking (18). Enzymatic reaction was terminated by adding 4× Laemmli sample buffer (Bio-Rad Laboratories, Hercules, CA, USA) with 10% β-mercaptoethanol (Sigma Aldrich, Saint Louis, MO, USA). AF samples were subsequently subjected to gel electrophoresis and analyzed by Western blotting as described below.

Janik K., Smith G.M, & Krynska B. (2023). Identification of Neurocan and Phosphacan as Early Biomarkers for Open Neural Tube Defects. Cells, 12(7), 1084.

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Enzymatic Tissue Digestion Protocol

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Explants from each animal (N = 6) were randomly assigned to an enzyme treatment group or the control culture media group (Fig. 1C). Explants were cultured in control (media) or treatment (media + enzyme) for 44hrs at 37 °C with media changes every 24hrs. Treatment groups were (1) 100 U/mL purified collagenase (Worthington Biochem Corp., NJ, USA, LS005273), (2) 50 U/mL bacterial hyaluronidase (MP Biomedicals, USA, 320421), or (3) 0.1 U/mL chondroitinase ABC (chABC) (Sigma Aldrich, MO, USA, C2905) (Sigma, USA). The concentrations used here were based on previous studies (Chan et al., 2010 (link); June and Fyhrie, 2009 (link); Karhula et al., 2017 (link); Nissi et al., 2016 (link); Pastrama et al., 2019 (link)). Samples were rinsed 3 × with PBS and enzyme inhibitors (5 mM EDTA and 5 mM benzamidine HCL), then stored in PBS on ice until testing.

McCreery K.P., Luetkemeyer C.M., Calve S, & Neu C.P. (2022). Hyperelastic characterization reveals proteoglycans drive the nanoscale strain-stiffening response in hyaline cartilage. Journal of biomechanics, 146, 111397.

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Modulating CSPG in Cell Culture

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Inhibition of CSPG through digestion of its CS chains was carried out by adding 50 mU/ml Chondroitinase ABC (ChABC; Sigma-Aldrich) diluted in 0.01% BSA (Sigma-Aldrich). Addition of external CSPG was achieved using 50 mg/ml proteoglycan from bovine nasal septum (Sigma-Aldrich), diluted in molecular grade water. Both treatments were added to the culture media every 48 h. As controls, cells were treated similarly with 0.01% BSA or water.

Hutchings C., Nuriel Y., Lazar D., Kohl A., Muir E., Genin O., Cinnamon Y., Benyamini H., Nevo Y, & Sela-Donenfeld D. (2024). Hindbrain boundaries as niches of neural progenitor and stem cells regulated by the extracellular matrix proteoglycan chondroitin sulphate. Development (Cambridge, England), 151(4), dev201934.

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CCL Proteoglycan Digestion Protocol

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The CCLs from both groups were immersed for 1 h at room temperature (20°C) in 20 ml buffer solution (15 ml of 20 mM Tris pH 7.5, 150 mM NaCl, 5 mM CaCl2) with protease inhibitors (1 tablet of mini-cOmplete per 10 ml of buffer, SIGMA-ALDRICH/Roche, United States) (Lujan et al., 2009 (link)). Chondroitinase ABC (ChABC) 0.25 IU/ml (SIGMA-ALDRICH, United States) enzyme was dissolved in 0.01% bovine serum albumin (BSA), and samples were incubated in this solution for 3 h prior to the mechanical tests as previously described (Lujan et al., 2009 (link)). This process was performed to reduce PG contents, and it was based on our preliminary work. Our preliminary work showed that approximately 80% of PGs in sectioned CCLs were digested within 3 h of incubation [Supplementary Materials (Supplementary Table S1 and Supplementary Figure S1)]. Control and treated groups were preserved during the mechanical tests in a custom-built tank filled with 600 ml of the buffer solution and protease inhibitors at room temperature (20°C) (1 tablet of cOmplete Protease Inhibitor Cocktail per 50 ml of buffer, SIGMA-ALDRICH/Roche, United States).

Readioff R., Geraghty B., Kharaz Y.A., Elsheikh A, & Comerford E. (2022). Proteoglycans play a role in the viscoelastic behaviour of the canine cranial cruciate ligament. Frontiers in Bioengineering and Biotechnology, 10, 984224.

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