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collagenase fractions  (PromoCell)


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    Structured Review

    PromoCell collagenase fractions
    Workflow for in Vitro ExtraCellular Matrix Mass Spectrometry Imaging (ivECM-MSI). 1.) Cells are seeded on Nunc Lab-Tek II glass chamber slides. Gelatin can be used as a coating prior to seeding if certain cell lines are not adhering to the glass. 2.) Cells are incubated in the glass chambers slides until confluence. 3.) The medium is removed and each well in the chamber slide is then decellularized with a 20 mM ammonium hydroxide solution for 5 min. Each well is then rinsed 4 times with sterile water. Next, the chambers are removed from the slide. Afterword, the slide is dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 4.) A 0.1 mg/mL solution of <t>collagenase</t> III is sprayed evenly onto the slides. The collagenase sprayed slides are then incubated in a 37 °C humidity chamber for 5 h to digest the collagen into peptides. Afterword, the slides are dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 5.) The collagenase digested slides are then sprayed evenly with a 7 mg/mL α-cyano-4-hydroxycinnamic acid (CHCA) solution. Immediately after CHCA spraying, a 5 mM ammonium phosphate solution is sprayed on top of the MALDI matrix to minimize matrix cluster formation. Slides at this stage can be temporarily stored in a vacuum desiccator if needed. 6.) The matrix sprayed slides are loaded into the MALDI mass spectrometer for MALDI mass spectrometry analysis. 7.) Finally, the data is visualized through mass spectrometry imaging heatmaps, m / z features are searched through a collagen peptide database, and statistics are performed. Created with BioRender.com .
    Collagenase Fractions, supplied by PromoCell, used in various techniques. Bioz Stars score: 97/100, based on 210 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase fractions/product/PromoCell
    Average 97 stars, based on 210 article reviews
    collagenase fractions - by Bioz Stars, 2026-03
    97/100 stars

    Images

    1) Product Images from "Profiling of collagen and extracellular matrix deposition from cell culture using in vitro ExtraCellular matrix mass spectrometry imaging (ivECM-MSI)"

    Article Title: Profiling of collagen and extracellular matrix deposition from cell culture using in vitro ExtraCellular matrix mass spectrometry imaging (ivECM-MSI)

    Journal: Matrix Biology Plus

    doi: 10.1016/j.mbplus.2024.100161

    Workflow for in Vitro ExtraCellular Matrix Mass Spectrometry Imaging (ivECM-MSI). 1.) Cells are seeded on Nunc Lab-Tek II glass chamber slides. Gelatin can be used as a coating prior to seeding if certain cell lines are not adhering to the glass. 2.) Cells are incubated in the glass chambers slides until confluence. 3.) The medium is removed and each well in the chamber slide is then decellularized with a 20 mM ammonium hydroxide solution for 5 min. Each well is then rinsed 4 times with sterile water. Next, the chambers are removed from the slide. Afterword, the slide is dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 4.) A 0.1 mg/mL solution of collagenase III is sprayed evenly onto the slides. The collagenase sprayed slides are then incubated in a 37 °C humidity chamber for 5 h to digest the collagen into peptides. Afterword, the slides are dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 5.) The collagenase digested slides are then sprayed evenly with a 7 mg/mL α-cyano-4-hydroxycinnamic acid (CHCA) solution. Immediately after CHCA spraying, a 5 mM ammonium phosphate solution is sprayed on top of the MALDI matrix to minimize matrix cluster formation. Slides at this stage can be temporarily stored in a vacuum desiccator if needed. 6.) The matrix sprayed slides are loaded into the MALDI mass spectrometer for MALDI mass spectrometry analysis. 7.) Finally, the data is visualized through mass spectrometry imaging heatmaps, m / z features are searched through a collagen peptide database, and statistics are performed. Created with BioRender.com .
    Figure Legend Snippet: Workflow for in Vitro ExtraCellular Matrix Mass Spectrometry Imaging (ivECM-MSI). 1.) Cells are seeded on Nunc Lab-Tek II glass chamber slides. Gelatin can be used as a coating prior to seeding if certain cell lines are not adhering to the glass. 2.) Cells are incubated in the glass chambers slides until confluence. 3.) The medium is removed and each well in the chamber slide is then decellularized with a 20 mM ammonium hydroxide solution for 5 min. Each well is then rinsed 4 times with sterile water. Next, the chambers are removed from the slide. Afterword, the slide is dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 4.) A 0.1 mg/mL solution of collagenase III is sprayed evenly onto the slides. The collagenase sprayed slides are then incubated in a 37 °C humidity chamber for 5 h to digest the collagen into peptides. Afterword, the slides are dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 5.) The collagenase digested slides are then sprayed evenly with a 7 mg/mL α-cyano-4-hydroxycinnamic acid (CHCA) solution. Immediately after CHCA spraying, a 5 mM ammonium phosphate solution is sprayed on top of the MALDI matrix to minimize matrix cluster formation. Slides at this stage can be temporarily stored in a vacuum desiccator if needed. 6.) The matrix sprayed slides are loaded into the MALDI mass spectrometer for MALDI mass spectrometry analysis. 7.) Finally, the data is visualized through mass spectrometry imaging heatmaps, m / z features are searched through a collagen peptide database, and statistics are performed. Created with BioRender.com .

    Techniques Used: In Vitro, Mass Spectrometry, Imaging, Incubation, Sterility



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    Workflow for in Vitro ExtraCellular Matrix Mass Spectrometry Imaging (ivECM-MSI). 1.) Cells are seeded on Nunc Lab-Tek II glass chamber slides. Gelatin can be used as a coating prior to seeding if certain cell lines are not adhering to the glass. 2.) Cells are incubated in the glass chambers slides until confluence. 3.) The medium is removed and each well in the chamber slide is then decellularized with a 20 mM ammonium hydroxide solution for 5 min. Each well is then rinsed 4 times with sterile water. Next, the chambers are removed from the slide. Afterword, the slide is dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 4.) A 0.1 mg/mL solution of <t>collagenase</t> III is sprayed evenly onto the slides. The collagenase sprayed slides are then incubated in a 37 °C humidity chamber for 5 h to digest the collagen into peptides. Afterword, the slides are dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 5.) The collagenase digested slides are then sprayed evenly with a 7 mg/mL α-cyano-4-hydroxycinnamic acid (CHCA) solution. Immediately after CHCA spraying, a 5 mM ammonium phosphate solution is sprayed on top of the MALDI matrix to minimize matrix cluster formation. Slides at this stage can be temporarily stored in a vacuum desiccator if needed. 6.) The matrix sprayed slides are loaded into the MALDI mass spectrometer for MALDI mass spectrometry analysis. 7.) Finally, the data is visualized through mass spectrometry imaging heatmaps, m / z features are searched through a collagen peptide database, and statistics are performed. Created with BioRender.com .
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    Image Search Results


    Workflow for in Vitro ExtraCellular Matrix Mass Spectrometry Imaging (ivECM-MSI). 1.) Cells are seeded on Nunc Lab-Tek II glass chamber slides. Gelatin can be used as a coating prior to seeding if certain cell lines are not adhering to the glass. 2.) Cells are incubated in the glass chambers slides until confluence. 3.) The medium is removed and each well in the chamber slide is then decellularized with a 20 mM ammonium hydroxide solution for 5 min. Each well is then rinsed 4 times with sterile water. Next, the chambers are removed from the slide. Afterword, the slide is dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 4.) A 0.1 mg/mL solution of collagenase III is sprayed evenly onto the slides. The collagenase sprayed slides are then incubated in a 37 °C humidity chamber for 5 h to digest the collagen into peptides. Afterword, the slides are dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 5.) The collagenase digested slides are then sprayed evenly with a 7 mg/mL α-cyano-4-hydroxycinnamic acid (CHCA) solution. Immediately after CHCA spraying, a 5 mM ammonium phosphate solution is sprayed on top of the MALDI matrix to minimize matrix cluster formation. Slides at this stage can be temporarily stored in a vacuum desiccator if needed. 6.) The matrix sprayed slides are loaded into the MALDI mass spectrometer for MALDI mass spectrometry analysis. 7.) Finally, the data is visualized through mass spectrometry imaging heatmaps, m / z features are searched through a collagen peptide database, and statistics are performed. Created with BioRender.com .

    Journal: Matrix Biology Plus

    Article Title: Profiling of collagen and extracellular matrix deposition from cell culture using in vitro ExtraCellular matrix mass spectrometry imaging (ivECM-MSI)

    doi: 10.1016/j.mbplus.2024.100161

    Figure Lengend Snippet: Workflow for in Vitro ExtraCellular Matrix Mass Spectrometry Imaging (ivECM-MSI). 1.) Cells are seeded on Nunc Lab-Tek II glass chamber slides. Gelatin can be used as a coating prior to seeding if certain cell lines are not adhering to the glass. 2.) Cells are incubated in the glass chambers slides until confluence. 3.) The medium is removed and each well in the chamber slide is then decellularized with a 20 mM ammonium hydroxide solution for 5 min. Each well is then rinsed 4 times with sterile water. Next, the chambers are removed from the slide. Afterword, the slide is dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 4.) A 0.1 mg/mL solution of collagenase III is sprayed evenly onto the slides. The collagenase sprayed slides are then incubated in a 37 °C humidity chamber for 5 h to digest the collagen into peptides. Afterword, the slides are dried in a vacuum desiccator. Slides can be stored in a −20 °C freezer for temporary storage if needed. 5.) The collagenase digested slides are then sprayed evenly with a 7 mg/mL α-cyano-4-hydroxycinnamic acid (CHCA) solution. Immediately after CHCA spraying, a 5 mM ammonium phosphate solution is sprayed on top of the MALDI matrix to minimize matrix cluster formation. Slides at this stage can be temporarily stored in a vacuum desiccator if needed. 6.) The matrix sprayed slides are loaded into the MALDI mass spectrometer for MALDI mass spectrometry analysis. 7.) Finally, the data is visualized through mass spectrometry imaging heatmaps, m / z features are searched through a collagen peptide database, and statistics are performed. Created with BioRender.com .

    Article Snippet: Pooled collagenase fractions were placed in Fibroblast Growth Medium 2 (Promocell, C-23020) with 10 % FBS and 1 % antibiotics/antimycotics to halt collagenase digestion, and then subjected to centrifugation at 1000 rpm for 5 mins at room temperature.

    Techniques: In Vitro, Mass Spectrometry, Imaging, Incubation, Sterility