fluorogenic adam10 substrate (Biozyme Laboratories)

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Biozyme Laboratories fluorogenic adam10 substrate
Fluorogenic Adam10 Substrate, supplied by Biozyme Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/fluorogenic+assay/adam10+fluorogenic+substrate/pm42006582-140-55-59
Average 86 stars, based on 1 article reviews

fluorogenic adam10 substrate - by Bioz Stars, 2026-11

86/100 stars

Images

Isolation:

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. Isolated cortical culture-derived sEVs were incubated at 37◦C with a fluorogenic ADAM10 substrate (‘PEPDAB063’, BioZyme, Inc.) in the presence or absence of ADAM10 inhibitor GI254023X (GI). (A) Schematic representation of the fluorogenic substrate assay, where ADAM10-mediated cleavage of the peptide separates the quencher (Dabcyl) from the fluorophore (5-FAM), resulting in increased fluorescence emission (~530 nm). (B) sEV-ADAM10 activity measurement was performed at 0.5, 3, and 24 h, with or without GI. ..

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. As protease activity is the result of a finely tuned interplay of many different factors and the mere presence of mature ADAM10 on EVs does not per se guarantee its activity there (Aljohmani et al., 2022 ; Lambrecht et al., 2018 ), we performed cell-free activity assays with the purified sEVs using a synthetic and fluorogenic ADAM10 substrate (PEPDAB063, BioZyme, Inc.) in the presence or absence of the selective ADAM10 inhibitor GI254023X (GI) (Figure 2A ). sEVs were isolated from cultured cortical cells as before but in the absence of a PI cocktail in the cell supernatants. ..

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: Images were acquired at 200 kV with a JEM2100Plus transmission electron microscope (Jeol) equipped with an XAROSA CMOS camera (Emsis). .. The proteolytic activity of sEV-ADAM10 was analysed using the ADAM10-selective fluorogenic substrate PEPDAB063 (Acetyl-dArg(3)-dGlu(3)-hexaminoyl-K(Dabcyl)PRYEAYKMGK(5FAM)-NH2) from BioZyme, Inc. (Caescu et al., 2009 ). sEVs were isolated from primary rat cortical cells as described above but without the use of PIs in the cell supernatant. .. Immediately after sEV isolation, sEVs were resuspended in cold 20 mM HEPES buffer (pH 7.0–7.6), split into equal parts by volume, and pre-incubated for 30 min with either the ADAM10-selective hydroxamate-based inhibitor GI254023X (GI) (Ludwig et al., 2005 ) (Selleck Chemicals) in DMSO (5 μM final concentration) or a control containing the same concentration of DMSO in 20 mM HEPES. sEV samples were transferred to black, flat-bottom, non-binding 96-well plates, and fluorogenic PEPDAB063 was added simultaneously to all wells (10 μM final concentration).

Incubation:

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. Isolated cortical culture-derived sEVs were incubated at 37◦C with a fluorogenic ADAM10 substrate (‘PEPDAB063’, BioZyme, Inc.) in the presence or absence of ADAM10 inhibitor GI254023X (GI). (A) Schematic representation of the fluorogenic substrate assay, where ADAM10-mediated cleavage of the peptide separates the quencher (Dabcyl) from the fluorophore (5-FAM), resulting in increased fluorescence emission (~530 nm). (B) sEV-ADAM10 activity measurement was performed at 0.5, 3, and 24 h, with or without GI. ..

Fluorescence:

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. Isolated cortical culture-derived sEVs were incubated at 37◦C with a fluorogenic ADAM10 substrate (‘PEPDAB063’, BioZyme, Inc.) in the presence or absence of ADAM10 inhibitor GI254023X (GI). (A) Schematic representation of the fluorogenic substrate assay, where ADAM10-mediated cleavage of the peptide separates the quencher (Dabcyl) from the fluorophore (5-FAM), resulting in increased fluorescence emission (~530 nm). (B) sEV-ADAM10 activity measurement was performed at 0.5, 3, and 24 h, with or without GI. ..

Activity Assay:

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. Isolated cortical culture-derived sEVs were incubated at 37◦C with a fluorogenic ADAM10 substrate (‘PEPDAB063’, BioZyme, Inc.) in the presence or absence of ADAM10 inhibitor GI254023X (GI). (A) Schematic representation of the fluorogenic substrate assay, where ADAM10-mediated cleavage of the peptide separates the quencher (Dabcyl) from the fluorophore (5-FAM), resulting in increased fluorescence emission (~530 nm). (B) sEV-ADAM10 activity measurement was performed at 0.5, 3, and 24 h, with or without GI. ..

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. As protease activity is the result of a finely tuned interplay of many different factors and the mere presence of mature ADAM10 on EVs does not per se guarantee its activity there (Aljohmani et al., 2022 ; Lambrecht et al., 2018 ), we performed cell-free activity assays with the purified sEVs using a synthetic and fluorogenic ADAM10 substrate (PEPDAB063, BioZyme, Inc.) in the presence or absence of the selective ADAM10 inhibitor GI254023X (GI) (Figure 2A ). sEVs were isolated from cultured cortical cells as before but in the absence of a PI cocktail in the cell supernatants. ..

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: Images were acquired at 200 kV with a JEM2100Plus transmission electron microscope (Jeol) equipped with an XAROSA CMOS camera (Emsis). .. The proteolytic activity of sEV-ADAM10 was analysed using the ADAM10-selective fluorogenic substrate PEPDAB063 (Acetyl-dArg(3)-dGlu(3)-hexaminoyl-K(Dabcyl)PRYEAYKMGK(5FAM)-NH2) from BioZyme, Inc. (Caescu et al., 2009 ). sEVs were isolated from primary rat cortical cells as described above but without the use of PIs in the cell supernatant. .. Immediately after sEV isolation, sEVs were resuspended in cold 20 mM HEPES buffer (pH 7.0–7.6), split into equal parts by volume, and pre-incubated for 30 min with either the ADAM10-selective hydroxamate-based inhibitor GI254023X (GI) (Ludwig et al., 2005 ) (Selleck Chemicals) in DMSO (5 μM final concentration) or a control containing the same concentration of DMSO in 20 mM HEPES. sEV samples were transferred to black, flat-bottom, non-binding 96-well plates, and fluorogenic PEPDAB063 was added simultaneously to all wells (10 μM final concentration).

Purification:

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. As protease activity is the result of a finely tuned interplay of many different factors and the mere presence of mature ADAM10 on EVs does not per se guarantee its activity there (Aljohmani et al., 2022 ; Lambrecht et al., 2018 ), we performed cell-free activity assays with the purified sEVs using a synthetic and fluorogenic ADAM10 substrate (PEPDAB063, BioZyme, Inc.) in the presence or absence of the selective ADAM10 inhibitor GI254023X (GI) (Figure 2A ). sEVs were isolated from cultured cortical cells as before but in the absence of a PI cocktail in the cell supernatants. ..

Cell Culture:

Article Title: ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.
Article Snippet: .. As protease activity is the result of a finely tuned interplay of many different factors and the mere presence of mature ADAM10 on EVs does not per se guarantee its activity there (Aljohmani et al., 2022 ; Lambrecht et al., 2018 ), we performed cell-free activity assays with the purified sEVs using a synthetic and fluorogenic ADAM10 substrate (PEPDAB063, BioZyme, Inc.) in the presence or absence of the selective ADAM10 inhibitor GI254023X (GI) (Figure 2A ). sEVs were isolated from cultured cortical cells as before but in the absence of a PI cocktail in the cell supernatants. ..

Review