Hyaluronidase 5

Manufactured by Merck Group

Hyaluronidase V is an enzyme that catalyzes the hydrolysis of hyaluronic acid, a component of the extracellular matrix. It is used in research applications to study the role of hyaluronic acid in various biological processes.

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Lab products found in correlation

Dnase 1, by Merck Group (6 mentions) Collagenase 4, by Merck Group (6 mentions) Pharm lyse, by BD (4 mentions) Dispase 2, by Roche (2 mentions) Soybean trypsin inhibitor, by Worthington (1 mentions) Red cell lysis buffer, by Solarbio (1 mentions)

Spelling variants of this product

Hyaluronidase (1464 citations) Hyaluronidase type V (31 citations)

6 protocols using hyaluronidase 5

Isolation and Dissociation of Tumor-Derived CD11b+ Cells

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Tumors were isolated, minced in a petri dish on ice and then enzymatically dissociated in Hanks Balanced Salt Solution containing 0.5 mg/ml Collagenase IV (Sigma), 0.1 mg/ml Hyaluronidase V (Sigma) and 0.005 MU/ml DNAse I (Sigma) at 37 °C for 5–30 min. The duration of enzymatic treatment was optimized for greatest yield of live CD11b+ cells per tumor type. Cell suspensions were filtered through a 70 μm cell strainer. Red blood cells were solubilized with red cell lysis buffer (Pharm Lyse, BD Biosciences, San Jose, CA), and the resulting suspension was filtered through a cell strainer to produce a single cell suspension. Cells were washed one time with PBS prior to use in flow cytometry analysis or sorting.

Schmid M.C., Khan S.Q., Kaneda M.M., Pathria P., Shepard R., Louis T.L., Anand S., Woo G., Leem C., Faridi M.H., Geraghty T., Rajagopalan A., Gupta S., Ahmed M., Vazquez-Padron R.I., Cheresh D.A., Gupta V, & Varner J.A. (2018). Integrin CD11b activation drives anti-tumor innate immunity. Nature Communications, 9, 5379.

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Isolation and Dissociation of Tumor-Infiltrating Cells

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Tumors were isolated, minced in a petri dish on ice and then enzymatically dissociated in Hanks Balanced Salt Solution containing 0.5 mg/ml Collagenase IV (Sigma), 0.1 mg/ml Hyaluronidase V (Sigma), 0.6 U/ml Dispase II (Roche) and 0.005 MU/ml DNAse I (Sigma) at 37°C for 5–30 min. The duration of enzymatic treatment was optimized for greatest yield of live CD11b+ cells per tumor type. Cell suspensions were filtered through a 70μm cell strainer. Red blood cells were solubilized with red cell lysis buffer (Pharm Lyse, BD Biosciences, San Jose, CA), and the resulting suspension was filtered through a cell strainer to produce a single cell suspension. Cells were washed one time with PBS prior to use in flow cytometry analysis or magnetic bead purification.

Kaneda M.M., Messer K.S., Ralainirina N., Li H., Leem C., Gorjestani S., Woo G., Nguyen A.V., Figueiredo C.C., Foubert P., Schmid M.C., Pink M., Winkler D.G., Rausch M., Palombella V.J., Kutok J., McGovern K., Frazer K.A., Wu X., Karin M., Sasik R., Cohen E.E, & Varner J.A. (2016). PI3Kγ is a molecular switch that controls immune suppression. Nature, 539(7629), 437-442.

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Dissociation of Tumor Tissue for Flow Cytometry

Cited 1 time

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Single-cell suspensions from tumors were prepared as described previously (20 (link)). Briefly, tumors were isolated, minced, and incubated for 30 to 45 minutes at 37°C in a dissociation solution containing Hanks Balanced Salt Solution supplemented with 0.5 mg/mL collagenase IV (Sigma), 0.1 mg/mL hyaluronidase V (Sigma), and 0.005 MU/ml DNAse I (Sigma). The undigested tissues were removed by passing through 70 μm nylon mesh and centrifuged at 1,500 rpm for 5 minutes. The red blood cells were lysed using RBC lysis buffer (Pharm Lyse; BD Biosciences) and cells were prepared for flow cytometry.

Rohila D., Park I.H., Pham T.V., Weitz J., Hurtado de Mendoza T., Madheswaran S., Ishfaq M., Beaman C., Tapia E., Sun S., Patel J., Tamayo P., Lowy A.M, & Joshi S. (2023). Syk Inhibition Reprograms Tumor-Associated Macrophages and Overcomes Gemcitabine-Induced Immunosuppression in Pancreatic Ductal Adenocarcinoma. Cancer Research, 83(16), 2675-2689.

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Isolation and Single-Cell Suspension of Tumors

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Tumors were isolated, minced in a petri dish on ice and then enzymatically dissociated in Hanks Balanced Salt Solution with Ca2+ and Mg2+ containing 0.5 mg/ml Collagenase IV (Sigma), 0.1 mg/ml Hyaluronidase V(Sigma), 0.6 U/ml Dispase II(Roche), 0.005 MU/ml DNAse I(Sigma), and 0.2 mg/ml soybean trypsin inhibitor(Worthington Biochemical) at 37 °C for 15 min. Red blood cells were lysed with red blood cell lysis solution, and the resulting suspension was filtered through a 70 um cell strainer to produce a single-cell suspension. Cells were then washed one time with PBS prior to incubation with fluorescently conjugated antibodies for flow cytometry.

Schmid M.C., Kang S.W., Chen H., Paradise M., Ghebremedhin A., Kaneda M.M., Chin S.M., Do A., Watterson D.M, & Varner J.A. (2022). PI3Kγ stimulates a high molecular weight form of myosin light chain kinase to promote myeloid cell adhesion and tumor inflammation. Nature Communications, 13, 1768.

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Enzymatic Dissociation of LLC Tumors

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The LLC tumors were minced and then enzymatically dissociated in Hanks Balanced Salt Solution containing 1 mg/mL Collagenase IV (Sigma), 0.1 mg/mL Hyaluronidase V (Sigma) and 5 μU/mL DNAse I (Sigma) at 37°C for 30 min. Red blood cells were solubilized with red cell lysis buffer (Solarbio), and the resulting suspension was filtered through a 70 μm cell strainer again to produce a single cell suspension for flow cytometry analysis.

Zhao C.C., Han Q.J., Ying H.Y., Gu X.X., Yang N., Li L.Y, & Zhang Q.Z. (2022). TNFSF15 facilitates differentiation and polarization of macrophages toward M1 phenotype to inhibit tumor growth. Oncoimmunology, 11(1), 2032918.

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Isolation of Single Cells from Tumor Tissue

Cited 1 time

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Single cell suspensions from tumors were prepared as described before (20 (link)). Briefly, tumors were isolated, minced, and incubated for 30–45 min at 37°C in a dissociation solution containing Hanks Balanced Salt Solution supplemented with 0.5 mg/ml collagenase IV (Sigma), 0.1 mg/ml hyaluronidase V (Sigma), and 0.005 MU/ml DNAse I (Sigma). The undigested tissues were removed by passing through 70 μm nylon mesh and centrifuged at 1500 rpm for 5 min. The red blood cells were lysed using RBC lysis buffer (Pharm Lyse, BD Biosciences, San Jose, CA, USA) and cells were prepared for flow cytometry.

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