Liberase tl

Manufactured by Roche

Sourced in Switzerland, United States, Germany, United Kingdom

Liberase TL is a digestive enzyme mixture used in laboratory applications. It is designed to dissociate tissue samples into single cells or small cell clusters. The product consists of a blend of purified enzymes that effectively break down the extracellular matrix, allowing for the isolation of viable cells from various tissues.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Liberase (1194 citations) Liberase TL Research Grade (30 citations) Liberase TL purified enzyme blend (11 citations) Liberase TL enzyme (5 citations) Liberase TL grade (5 citations) Liberase TL solution (4 citations) Liberase TL Research Grade enzyme (3 citations) Liberase TL enzyme blend (3 citations) Liberase DL/Liberase TL andDNASEI (2 citations) Liberase™ TL enzyme preparations (2 citations)

505 protocols using liberase tl

Multi-tissue Immune Cell Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Inguinal and popliteal lymph nodes were removed and passed through a 70-µm cell strainer. Blood was collected together with heparin followed by two rounds of red blood cell lysis or collected without an anticoagulant and allowed to clot for serum collection. Cells from the paw were dissociated using 25 µg/ml of Liberase TL (Roche) and 1 mg/ml of DNaseI (Roche). Air pouch infiltrate was harvested 4 h after challenge by washing the pouch with PBS followed by centrifugation to separate cells from lavage. Air pouch membrane was carefully dissected and digested for 1 h with Liberase TL (Roche), and then passed through a cell strainer. Neutrophils were isolated from joints or the blood using CD11b MicroBeads (Miltenyi Biotec) followed by 2 h incubation at 37°C to separate adherent cells (macrophage-enriched) from nonadherent cells (neutrophil-enriched). Neutrophils were also purified from air pouch infiltrates or bone marrow using a Percoll gradient (52%/64%/72%) with 85–95% purity obtained in the bottom band (interface of 64%/72%). Infiltrating cells from the air pouch were stained for Annexin V and Propidium Iodide (eBioscience) uptake and analyzed by flow cytometry to determine the percent undergoing apoptosis.

Blazek K., Eames H.L., Weiss M., Byrne A.J., Perocheau D., Pease J.E., Doyle S., McCann F., Williams R.O, & Udalova I.A. (2015). IFN-λ resolves inflammation via suppression of neutrophil infiltration and IL-1β production. The Journal of Experimental Medicine, 212(6), 845-853.

+ Open protocol

+ Expand

3D Aggregation and Disaggregation of MSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

MSCs were seeded into U-bottomed 96-well plates with 30,000, 60,000, or 120,000 cells per well in medium containing 0.25% methyl cellulose. Cells self-aggregated to form 3D MSCs within 5 hours. For studies requiring disaggregation, 3D MSCs were collected, washed in PBS and then resuspended in Liberase TL working solution (Liberase TL (Roche) 1.3 Wunsch units per ml in PBS). 3D MSCs were incubated on an orbital shaker at 150 rpm for 20 minutes at 37 °C and then disaggregated to a single cell suspension by pipetting, before reseeding onto tissue culture plastic in 2D MSC media. Cell areas of in vitro-aged MSCs, before and after 3D culture, were calculated using Image J.

Pennock R., Bray E., Pryor P., James S., McKeegan P., Sturmey R, & Genever P. (2015). Human cell dedifferentiation in mesenchymal condensates through controlled autophagy. Scientific Reports, 5, 13113.

+ Open protocol

+ Expand

Extraction of Lymphocytes from Skin, Lung and Liver

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lymphocytes were extracted from the skin as previously described [21 (link)]. Briefly, a small piece of dorsal skin was removed, minced and incubated with serum free media containing Liberase TL (250 μg/ml; Roche) and 0.05% DNase I (Roche) for 105 min at 37°C. A single cell suspension was prepared by mashing the skin tissue slurry through a 100μm nylon mesh. Cells were sequentially filtered through 70 and 40μm nylon filters [21 (link)]. Lung and liver infiltrating lymphocytes were extracted as follows: lung and liver were removed, minced, and digested in serum free media containing Liberase TL (250μg/ml; Roche) and 0.05% DNase I (Roche) for 30 and 15min respectively. Lung and liver infiltrating lymphocytes were separated by centrifugation in 40% Percoll and collected in a cell pellet.

Amarnath S., Foley J.E., Farthing D.E., Gress R.E., Laurence A., Eckhaus M.A., Métais J.Y., Rose J.J., Hakim F.T., Felizardo T.C., Cheng A.V., Robey P.G., Stroncek D.E., Sabatino M., Battiwalla M., Ito S., Fowler D.H, & Barrett A.J. (2015). Bone Marrow Derived Mesenchymal Stromal Cells Harness Purinergenic Signaling to Tolerize Human Th1 Cells In Vivo. Stem cells (Dayton, Ohio), 33(4), 1200-1212.

+ Open protocol

+ Expand

Epidermal and Dermal Cell Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Epidermal and dermal cell suspensions were prepared from the trunk skin of TIM-4 WT and KO mice, as previously reported [72 (link), 73 (link)]. In brief, trunk skin was incubated in 0.5% dispase (Gibco, Life Technologies, Grand Island, NY, USA) for 1 hour at 37°C after its subcutaneous fat was scraped off. Then, epidermal sheet was peeled from the dermis, cut into tiny pieces and digested in complete culture medium containing 0.01% DNase (Sigma, St Louis, MO, USA) for 1 hour at 37°C. The underlying dermis was cut into approximately 3 × 3 mm pieces, which were floated in 0.04% Liberase TL (Roche Diagnostics GmbH, Mannheim, Germany) for 45 minutes and subsequently 0.04% Liberase TL + 0.005% DNase for additional 15 minutes at 37°C. The epidermal and dermal single-cell suspensions were harvested after filtering through a 70 μM filter. Complete culture medium was RPMI 1640 (with 2mM L-glutamine, Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Hyclone, Thermo Scientific, Pittsburgh, PA, USA), 5 × 10 ⁵ M 2-mercaptoethanol, 0.15% sodium hydrogencarbonate, 1 mM sodium pyruvate, nonessential amino acids, 100U per ml penicillin and 0.01% streptomycin.

Zhang X., Liu Q., Wang J., Li G., Weiland M., Yu F.S., Mi Q.S., Gu J, & Zhou L. (2016). TIM-4 is differentially expressed in the distinct subsets of dendritic cells in skin and skin-draining lymph nodes and controls skin Langerhans cell homeostasis. Oncotarget, 7(25), 37498-37512.

+ Open protocol

+ Expand

Anti-CD137 Immunotherapy in Murine Tumor Model

Check if the same lab product or an alternative is used in the 5 most similar protocols

WT and Batf3^-/- mice were inoculated s.c. with 2 x 10⁶ MC38-OVA cells. Mice were injected i.p. with 100 µg anti-CD137 or an isotype control at days 5 and 7 after tumor inoculation. LNs and tumors were extracted at day 9. LNs were incubated at 37ºC in Liberase TL (Roche, 20 minutes) and tumors in Liberase TL/DNase I (30 minutes). Then, both LN and tumors were mechanically dissociated through a 70 µm cell strainer (Fisher Scientific). Single cell suspensions were stained and analyzed by flow cytometry.
For OVA- or Adpgk-specific T cell restimulation ex-vivo, single cell suspensions from LNs were cultured for 2 h in 10% FBS RPMI medium containing 1 µg/ml SIINFEKL or ASMTNMELM peptide. Then Brefeldin A was added at a final concentration of 10 µg/ml and cells were incubated for 10 h. Cells were stained for surface markers, fixed and permeabilized for intracellular IFN-γ staining. Samples were analyzed by flow cytometry.

Sánchez-Paulete A.R., Cueto F.J., Martínez-López M., Labiano S., Morales-Kastresana A., Rodríguez-Ruiz M.E., Jure-Kunkel M., Azpilikueta A., Aznar M.A., Quetglas J.I., Sancho D, & Melero I. (2015). Cancer immunotherapy with immunomodulatory anti-CD137 and anti-PD-1 monoclonal antibodies requires Batf3-dependent dendritic cells. Cancer discovery, 6(1), 71-79.

+ Open protocol

+ Expand

Isolation and Characterization of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lung, pancreas, kidney, liver and spleen were minced. Minced lung was incubated with a digestion cocktail containing 400 μg/ml Liberase TL (Roche) in IMDM medium for 30 minutes at 37°C with shaking. Lung, pancreas, kidney, liver, spleen, lymph nodes, thymus and Payer’s patches were mashed through a 70μl cell strainer to obtain single cell suspensions. Cells from skin were isolated as described above.
Gut-resident lymphocytes were prepared by cutting the small intestine or large intestine (colon and cecum) in small pieces and incubation in digestion buffer (IMDM supplemented with 3% (vol/vol) FCS, 5 mM EDTA, 25 mM HEPES, penicillin/streptomycin and 0.145mg/ml DTT for 20-40 min at 37°C, shaking. The supernatant containing epithelial and intraepithelial cells (IEL) was enriched for IEL using 36.5% Percoll (Amersham) density gradient centrifugation. The remaining tissue was further digested with a cocktail containing 400 μg/ml Liberase TL (Roche) and DNase I (10 U/ml; Sigma) for 30 min at 37 °C. For flow cytometry analysis cells were stained for surface molecules (see list of antibodies in Table 1) in the presence of 10% Fc block and acquired on a FACSCantoII (BD).

Ahlfors H., Morrison P.J., Duarte J.H., Li Y., Biro J., Tolaini M., Di Meglio P., Potocnik A.J, & Stockinger B. (2014). IL-22 fate reporter reveals origin and control of IL-22 production in homeostasis and infection. Journal of immunology (Baltimore, Md. : 1950), 193(9), 4602-4613.

+ Open protocol

+ Expand

Tissue Dissociation for Single-Cell Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cohort A tissue was vigorously minced with surgical scissors and transferred to a GenleMACs™ C Tube (Miltenyi Biotec) with 100 μg/mL Liberase TL (Roche) and 200 μg/mL DNase I (Roche) at 3mL per gram of tissue. C Tubes were incubated in the GentleMACS™ octo dissociator (Miltenyi Biotec) with heaters following the manufacturers dissociation protocol (Miltenyi Biotec Tumor Dissociation Kit). 10mL of sort buffer (PBS + 2% fetal calf Serum (FCS) + 2mM EDTA) was added to samples and filtered through a 100 μm filter, spun down, and red blood cells lysed with 175mM ammonium chloride. Cohort B tissue was vigorously minced with surgical scissors and transferred to a 50 mL conical with 20 μL/mL Liberase TL (at 5 mg/mL, Roche) and 50 U/ml DNase I (Roche) per 0.3 g of tissue for 30 minutes at 37° C with 5 % CO₂ with constant agitation. Samples were then filtered through a 70 μm filter, spun down, and re-suspended for staining³¹.

Barry K.C., Hsu J., Broz M.L., Cueto F.J., Binnewies M., Combes A.J., Nelson A.E., Loo K., Kumar R., Rosenblum M.D., Alvarado M.D., Wolf D.M., Bogunovic D., Bhardwaj N., Daud A.I., Ha P.K., Ryan W.R., Pollack J.L., Samad B., Asthana S., Chan V, & Krummel M.F. (2018). A Natural Killer/Dendritic Cell Axis Defines Responsive Tumor Microenvironments in Melanoma. Nature medicine, 24(8), 1178-1191.

+ Open protocol

+ Expand

Mouse Pancreatic Islet Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

The mouse pancreas was first perfused with 0.125 mg/ml LiberaseTL (Roche, Nutley, NJ, USA) from the biliary-pancreatic bile duct, then was incubated in 0.125 mg/ml LiberaseTL in a 37 o C shaker at 200rpm for 45 minutes. Histopaque1077 and Histopaque1119 (Sigma-Aldrich, USA) were mixed at a ratio of 9:33 (vol:vol) to generate Histopaque1110. Centrifugation in Histopaque1100 at 1200rpm for 15 minutes without brake (islets are in the solution fraction), following by hand-pickings, was then performed to isolate islets.

Wang C., Chen X., Ding X., He Y., Gu C, & Zhou L. (2015). Exendin-4 Promotes Beta Cell Proliferation via PI3k/Akt Signalling Pathway. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 35(6).

+ Open protocol

+ Expand

Isolation and Culture of Mouse DRG Neurons

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

All experimental procedures were approved by the Institutional Animal Care and Use Committee of Johns Hopkins University School of Medicine and were in accordance with the guidelines provided by the National Institute of Health and the International Association for the Study of Pain. Isolation of dorsal root ganglion (DRG) neurons and primary culture were conducted as described^{60 (link)}. Briefly, DRG were isolated from 6–12 week old C57Bl6 wild type mice, rinsed in Complete Saline Solution (CSS containing, in mM: 137 NaCl, 5.3 KCl, 1 MgCl₂, 3 CaCl₂, 25 sorbitol, and 10 mM HEPES pH 7.2) and digested with TM Liberase (0.35 U/ml Roche) in in CSS/0.5 mM EDTA at 37 °C for 20 min, followed by TL Liberase (0.25 U/ml, Roche) in CSS/0.5 mM EDTA supplemented with papain (30 U/ml, Worthington) for 15 min, centrifuged and resuspended in DMEM containing BSA (1 mg/ml, Sigma) and Trypsin Inhibitor (1 mg/ml, Sigma), and mechanically dissociated. Cells were passed through a 70 micron strainer and spotted on poly-l-lysine/laminin coated coverslips. After 1 hr. of adherence, complete DRG medium (DMEM/F12, 10% FBS, 1% glutamine, antibiotics), was added to each well.

Ostrow K.L., Donaldson K.J., Caterina M.J., Belzberg A, & Hoke A. (2019). The Secretomes of Painful Versus Nonpainful Human Schwannomatosis Tumor Cells Differentially Influence Sensory Neuron Gene Expression and Sensitivity. Scientific Reports, 9, 13098.

+ Open protocol

+ Expand

Isolation of Skin Cell Types

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were sacrificed, shaved, and the non-wounded back skin or wound biopsies were harvested. In case of normal back skin, the subcutaneous fat was scraped off, and epidermis and dermis were separated using 0.25% dispase (Gibco, Zug, Switzerland) for 50min at 37°C under continuous shaking. The epidermis was treated with 100 KUnits/ml DNase (Sigma) in RPMI supplemented with penicillin/streptomycin (P/S), 10% fetal bovine serum and 20mM HEPES for 45 min at 37°C under continuous shaking. The dermis as well as the wound biopsies were cut into small pieces and treated with 0.25mg/ml TL Liberase (Roche) in RPMI supplemented with P/S and 20 mM HEPES for 1h at 37°C under continuous shaking. Finally, single cell suspensions were prepared by straining the mixture through 70 μm cell strainers, and cells were washed with 1x PBS.

Joshi N, & Werner S. (2017). Nrf2 is highly expressed in neutrophils, but myeloid cell-derived Nrf2 is dispensable for wound healing in mice. PLoS ONE, 12(10), e0187162.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!