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Liberase 2

Manufactured by Roche
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Liberase 2 is a laboratory product designed for tissue dissociation. It is a mixture of enzymes that effectively breaks down extracellular matrices, facilitating the isolation of cells from tissue samples.

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Lab products found in correlation

Egm2 2 fbs, by Lonza (2 mentions) Anti cd31 conjugated beads, by Miltenyi Biotec (2 mentions) Anti cd34 beads, by Miltenyi Biotec (2 mentions) Trypsin edta, by Thermo Fisher Scientific (2 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Magnetic activated cell sorting (macs), by Miltenyi Biotec (1 mentions) Egm 2, by Lonza (1 mentions) Huvecs, by Lonza (1 mentions)

Close spelling variants of this product

Liberase II kit Liberase type II Liberase Blendzyme 2 Liberase

6 protocols using liberase 2

1

Isolation and Culture of Cardiac SVPs

Cited 2 times
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SVPs were obtained as previously described by digestion of vein leftover, obtained from cardiac bypass patients with their signed consent, as approved by the Bath Research Ethics Committee (approval number 06/Q2001/1970).[10 (link)] Briefly, venous samples were minced and digested with Liberase 2 (Roche, Basel, Switzerland). SVPs were derived from CD34positive/CD31negative cells isolated by magnetic bead–assisted cell sorting (MACS, Miltenyi).[10 (link),12 (link)]
Both human umbilical vein endothelial cells (HUVECs, Lonza) and SVPs were cultured in endothelial growth medium (EGM2, Lonza); SVPs were seeded on gelatin and fibronectin coated plates, as previously described.[10 (link),12 (link)]
Campagnolo P., Gormley A.J., Chow L.W., Guex A.G., Parmar P.A., Puetzer J.L., Steele J.A., Breant A., Madeddu P, & Stevens M.M. (2016). Pericyte seeded dual peptide scaffold with improved endothelialization for vascular graft tissue engineering. Advanced healthcare materials, 5(23), 3046-3055.
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2

Isolation of Pericyte Progenitor Cells

Cited 1 time
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Pericyte progenitor cells were isolated from vein leftovers of patients undergoing coronary artery bypass graft surgery or varicose vein removal, as previously described 55 (link). In brief, saphenous veins were carefully dissected from surrounding tissues using a sterile scalpel and then thoroughly washed in PBS. Veins were manually minced prior to 4 hours incubation with 3.7 mg/mL Liberase 2 (Roche) and filtered passing the cell suspension through 30μm cell strainer. Cells were depleted for endothelial cells with anti-CD31 conjugated beads (Miltenyi), according to manufacturer’s instruction. The remaining cells were purified by selecting CD34+ cells by anti-CD34 beads (Miltenyi). Target cells were then plated on fibronectin (10 μg/mL) coated plates in presence of differentiation medium (EGM2-2% FBS, Lonza). Adherent colonies were passaged to new culture dishes once they reached 60-70% confluence and frozen stocks generated after Passage 2 (P2) for the experiments shown in this publication. Trypsin-EDTA (Life Technologies) was utilized to detach cells from the growth substrate.
Caporali A., Meloni M., Nailor A., Mitić T., Shantikumar S., Riu F., Sala-Newby G.B., Rose L., Besnier M., Katare R., Voellenkle C., Verkade P., Martelli F., Madeddu P, & Emanueli C. (2015). p75NTR-dependent activation of NF-κB regulates microRNA-503 transcription and pericyte–endothelial crosstalk in diabetes after limb ischaemia. Nature Communications, 6, 8024.
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3

Isolation of Pericyte Progenitor Cells

Cited 1 time
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Pericyte progenitor cells were isolated from vein leftovers of patients undergoing coronary artery bypass graft surgery or varicose vein removal, as previously described55 (link). In brief, saphenous veins were carefully dissected from surrounding tissues using a sterile scalpel and then thoroughly washed in PBS. Veins were manually minced before 4-h incubation with 3.7 mg ml−1 Liberase 2 (Roche) and filtered passing the cell suspension through a 30-μm cell strainer. Cells were depleted for ECs with anti-CD31-conjugated beads (Miltenyi), according to the manufacturer's instruction. The remaining cells were purified by selecting CD34+ cells by anti-CD34 beads (Miltenyi). Target cells were then plated on fibronectin (10 μg ml−1)-coated plates in presence of differentiation medium (EGM-2—2% FBS, Lonza). Adherent colonies were passaged to new culture dishes once they reached 60–70% confluence and frozen stocks generated after Passage 2 (P2) for the experiments shown in this publication. Trypsin-EDTA (Life Technologies) was utilized to detach cells from the growth substrate.
Caporali A., Meloni M., Nailor A., Mitić T., Shantikumar S., Riu F., Sala-Newby G.B., Rose L., Besnier M., Katare R., Voellenkle C., Verkade P., Martelli F., Madeddu P, & Emanueli C. (2015). p75NTR-dependent activation of NF-κB regulates microRNA-503 transcription and pericyte–endothelial crosstalk in diabetes after limb ischaemia. Nature Communications, 6, 8024.
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4

Isolation and Purification of Murine Islets

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Murine islets were isolated from age- and sex-matched control and miR-206KO mice, as described (31 (link)). Briefly, mice were euthanized by cervical dislocation. The common bile duct was cannulated using a 27G needle, and 2 ml of ice-cold Liberase II (Roche Diagnostics, Basel, Switzerland) was injected immediately. The perfused pancreas was dislodged from the intestine, spleen, and stomach. To facilitate complete digestion, the pancreas was further incubated in a water bath preheated at 37°C for predetermined time intervals (batch variation, average time 12 min and 30 s). After digestion, islets were purified by filtration and gradient separation. Islets were maintained in RPMI (GIBCO; Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum and 100 U/ml penicillin-streptomycin (GIBCO) for 24 h, and healthy islets were hand-picked for experiments.
Vinod M., Patankar J.V., Sachdev V., Frank S., Graier W.F., Kratky D, & Kostner G.M. (2016). MiR-206 is expressed in pancreatic islets and regulates glucokinase activity. American journal of physiology. Endocrinology and metabolism, 311(1), E175-E185.
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5

Isolation and Purification of Murine Islets

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Murine islets were isolated from age- and sex-matched control and miR-206KO mice, as described (31 (link)). Briefly, mice were euthanized by cervical dislocation. The common bile duct was cannulated using a 27G needle, and 2 ml of ice-cold Liberase II (Roche Diagnostics, Basel, Switzerland) was injected immediately. The perfused pancreas was dislodged from the intestine, spleen, and stomach. To facilitate complete digestion, the pancreas was further incubated in a water bath preheated at 37°C for predetermined time intervals (batch variation, average time 12 min and 30 s). After digestion, islets were purified by filtration and gradient separation. Islets were maintained in RPMI (GIBCO; Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum and 100 U/ml penicillin-streptomycin (GIBCO) for 24 h, and healthy islets were hand-picked for experiments.
Vinod M., Patankar J.V., Sachdev V., Frank S., Graier W.F., Kratky D, & Kostner G.M. (2016). MiR-206 is expressed in pancreatic islets and regulates glucokinase activity. American journal of physiology. Endocrinology and metabolism, 311(1), E175-E185.
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6

Neutrophil Analysis in Bone Marrow Chimeras

Cited 1 time
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WT and Syk-deficient bone marrow chimeras were injected with control or pathogenic IgG. On days 8 through 10, mice were killed, and their ears were removed. The samples were digested with Liberase II (Roche, Basel, Switzerland) (Weber et al., 2015 (link)). Single-cell suspensions were obtained, and neutrophil numbers were determined by flow cytometry on the basis of their forward and side scatter characteristics and Ly6G-positivity (clone 1A8, BD Biosciences, San Jose, CA).
Németh T., Virtic O., Sitaru C, & Mócsai A. (2017). The Syk Tyrosine Kinase Is Required for Skin Inflammation in an In Vivo Mouse Model of Epidermolysis Bullosa Acquisita. The Journal of Investigative Dermatology, 137(10), 2131-2139.
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