Collagenase d and a

Manufactured by Roche

Collagenase D and A are enzymes used for dissociation and isolation of cells from tissues. Collagenase D and A function by breaking down collagen, a structural component of the extracellular matrix.

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Lab products found in correlation

Pharm lyse, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Facsaria 3 sorter, by BD (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) 40 μm cell strainer, by BD (1 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions) Fc block, by Thermo Fisher Scientific (1 mentions) Cd11b bv510, by BD (1 mentions) Cd64 pe, by BD (1 mentions) Ly6c apc, by BD (1 mentions) Cd45 pb, by BD (1 mentions) Ly6g percp cy5, by BD (1 mentions) Facscanto, by BD (1 mentions)

Close spelling variants of this product

Collagenase D Collagenase A Collagenase

3 protocols using collagenase d and a

Isolation and Analysis of Lung Immune Cells

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Mouse lungs perfused with PBS were minced and digested with Collagenase D and A (1 mg/mL each, Roche) in 2 mL for 1 hour at 37°C. Single cell suspension using 40μm cell strainers was prepared; erythrocytes were lysed with PharmLyse (BD Biosciences) (15 (link)). Cells were incubated with anti-mouse CD16/32 mAb and stained with directly-labeled antibodies against CD45, CD11b, F4/80, Ly6G, Ly6C and CD31 (BD Biosciences). Blood samples were treated with PharmLyse and stained as described above. Data were acquired on a FACS Canto II (BD Biosciences) and analyzed using Flow Jo software (Tree Star).

Häuselmann I., Roblek M., Protsyuk D., Huck V., Knopfova L., Grässle S., Bauer A.T., Schneider S.W, & Borsig L. (2016). Monocyte induction of E-selectin-mediated endothelial activation releases VE-cadherin junctions to promote tumor cell extravasation in the metastasis cascade. Cancer research, 76(18), 5302-5312.

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Isolation of Lung Endothelial Cells and Inflammatory Monocytes

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PBS-perfused lungs were minced and digested in Collagenase D and A (1 mg/mL each, Roche) for 1 hour at 37°C. A single-cell suspension was prepared by passing the digested lungs through 40μm cell strainers (BD Biosciences). Cells were incubated with antibodies against CD31, CD45, CD11b, Ly6C and Ly6G (eBiosciences). Endothelial cells (CD45^-CD11b^-CD31⁺) and inflammatory monocytes (CD45⁺CD11b⁺Ly6C^highLy6G^-) were sorted with a FACSAria III sorter (BD Biosciences). Total RNA was isolated using the RNeasy Mini Kit (Qiagen).

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Profiling Tumor-Associated Immune Cells

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The lungs of 4T1-tumor bearing mice were perfused, minced and digested in Collagenase D and A (2 mg each, Roche) for 1 hour at 37°C. A single-cell suspension using 40μm cell strainers was prepared, erythrocytes lysed. Cells were resuspended in staining buffer with F_c block (eBioscience), incubated on ice for 10 min, followed by staining with following antibodies: CD45-PB, CD11b-BV510, CD64-PE, Ly6C-APC, Ly6G-PerCP-Cy5.5 (all from BD) for 30 min. Data were collected with CountBright absolute counting beads (Life Technologies) using a FACSCanto (BD) and analyzed by FlowJo software (FlowJo, LCC).

Knopfová L., Biglieri E., Volodko N., Masařík M., Hermanová M., Garzón J.G., Dúcka M., Kučírková T., Souček K., Šmarda J., Beneš P, & Borsig L. (2017). Transcription factor c-Myb inhibits breast cancer lung metastasis by suppression of tumor cell seeding. Oncogene, 37(8), 1020-1030.

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