Collagenase 4

Manufactured by Worthington

Sourced in United States, Switzerland, China, Germany

Collagenase IV is an enzyme used in laboratory settings to break down collagen, a structural protein found in various tissues. It is commonly used to isolate cells from tissue samples for further research and experimentation.

Automatically generated - may contain errors

Lab products found in correlation

Dnase 1, by Roche (68 mentions) Dnase 1, by Merck Group (52 mentions) Dnase 1, by Worthington (35 mentions) Hyaluronidase, by Merck Group (28 mentions) Collagenase 2, by Worthington (18 mentions) Dnase, by Merck Group (18 mentions) Percoll, by GE Healthcare (16 mentions) Collagenase 1, by Worthington (15 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (14 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (13 mentions) Dispase, by Thermo Fisher Scientific (12 mentions) Gentlemacs dissociator, by Miltenyi Biotec (11 mentions) Trypsin edta, by Thermo Fisher Scientific (11 mentions) Dnase, by Worthington (10 mentions) Dnase type 4, by Merck Group (9 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (9 mentions) 70 μm cell strainer, by BD (9 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions) Collagenase d, by Roche (8 mentions) Gentlemacs tissue dissociator, by Miltenyi Biotec (8 mentions)

Close spelling variants of this product

Collagenases IV Collagenase I and IV Collagenase IV (LS004188) Collagenase IV (CLS-4, Collagenase I-IV Collagenase type I and IV Ultrapure collagenase IV Collagenase type II and IV Collagenase type IV solution Collagenase type IV

448 protocols using collagenase 4

Isolation of Splenic Stromal Cells

Cited 1 time

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

Splenic single-cell suspensions were prepared by mechanical disruption. The isolation of stromal cells from spleen has been described68 (link). Briefly, spleens were flushed with RPMI containing 3 mg ml⁻¹ Collagenase IV (Worthington), 40 μg ml⁻¹ DNAseI (Roche) and 2% (vol/vol) FCS), were cut into pieces and digested. Erythrocytes were lysed in 0.15 M NH₄Cl/10 mM KHCO₃/0.1 mM EDTA and haematopoietic CD45⁺ cells were depleted using anti-CD45 beads (Miltenyi Biotec). For TIL analysis, tumours were cut and digested with accutase (PAA), Collagenase IV (Worthington), Hyaluronidase (Sigma) and DNAseI (Roche) and red blood cells were lysed. Mononuclear cells were isolated using a Histopaque-1119 gradient.

Kallert S.M., Darbre S., Bonilla W.V., Kreutzfeldt M., Page N., Müller P., Kreuzaler M., Lu M., Favre S., Kreppel F., Löhning M., Luther S.A., Zippelius A., Merkler D, & Pinschewer D.D. (2017). Replicating viral vector platform exploits alarmin signals for potent CD8+ T cell-mediated tumour immunotherapy. Nature Communications, 8, 15327.

+ Open protocol

+ Expand

Isolation and Purification of Tumor-associated Immune Cells

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

Subcutaneous, orthotopic or MMTV-PyMT tumours were excised, cut in small pieces, treated with 10 U ml⁻¹ collagenase I, 400 U ml⁻¹ collagenase IV and 30 U ml⁻¹ DNaseI (Worthington) for 30 min at 37 °C, squashed and filtered. Red blood cells were removed using erythrocyte lysis buffer and density gradients (Axis-Shield) were used to remove debris and dead cells.
Tumour-draining LNs were cut, dissociated with 10 U ml⁻¹ collagenase I, 400 U ml⁻¹ collagenase IV and 30 U mL⁻¹ DNaseI (Worthington) for 45 min at 37 °C and filtered.
Spleens were flushed with 200 U ml⁻¹ collagenase III (Worthington) and left for 30 min at 37 °C. Afterwards, spleens were filtered and red blood cells were removed using erythrocyte lysis buffer.
To purify DC subpopulations from tumour, spleen or LNs, CD11c⁺ cells were MACS-enriched (anti-CD11c microbeads; Miltenyi) and sorted using BD FACSAria II (BD Biosciences) according to the gating strategy in Fig. 1a, Supplementary Fig. 6A or Supplementary Fig. 8A, respectively.
Bone marrow leukocytes were isolated through flushing of tibia and femur. The obtained cell suspensions were filtered, and red blood cells were removed using erythrocyte lysis buffer. To purify bone marrow monocytes, CD11b⁺ cells were MACS-enriched (anti-CD11b microbeads; Miltenyi) before sorting.

Laoui D., Keirsse J., Morias Y., Van Overmeire E., Geeraerts X., Elkrim Y., Kiss M., Bolli E., Lahmar Q., Sichien D., Serneels J., Scott C.L., Boon L., De Baetselier P., Mazzone M., Guilliams M, & Van Ginderachter J.A. (2016). The tumour microenvironment harbours ontogenically distinct dendritic cell populations with opposing effects on tumour immunity. Nature Communications, 7, 13720.

+ Open protocol

+ Expand

Establishment and Propagation of Patient-Derived Xenograft Models

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

Patient tumor samples to be used in PDX models were collected and processed by the University of Iowa College of Medicine Tissue Procurement Core under IRB# 200804792. Tissue collection and distribution was performed in accordance with the guidelines of the University of Iowa Institutional Review Board. Briefly, following collection tumor samples were placed in DMEM containing 5% FBS, 1X Pen/Strep, and 1% fungizone. For initiation of PDX tumors, patient tumor samples were processed by mincing until they passed through an 18 G needle easily. The cells were then suspended in a 1:1 mixture of DMEM (GIBCO) and Matrigel (Corning, 354230), and subcutaneously injected into 2 or 3 NOD.Cg-Prkdc^scid Il2^rgtmWjl/SzJ (NSG) (Jackson Laboratory) mice. For the propagation of samples, tumors were dissociated into single-cell suspensions and each mouse was injected subcutaneously with 1x10⁶ cells. Single-cell suspensions were generated by placement of the tumor into a sterile Petri dish containing 10 mL collagenase IV solution (9 mL HBSS (Ca²⁺- and Mg²⁺-free), 1 mL collagenase IV (Worthington Biochemical Corp.), and 5 mM CaCl₂). The tumor samples were first minced and incubated for 1 hr at 37°C. Following this, samples were further digested in 10 mL 0.05% trypsin (at 37°C for 10 min), and passed through an 18 G needle 30 times, after which the suspension was filtered using a 70 μm cell strainer.

Moose D.L., Krog B.L., Kim T.H., Zhao L., Williams-Perez S., Burke G., Rhodes L., Vanneste M., Breheny P., Milhem M., Stipp C.S., Rowat A.C, & Henry M.D. (2020). Cancer Cells Resist Mechanical Destruction in Circulation via RhoA/Actomyosin-Dependent Mechano-Adaptation. Cell reports, 30(11), 3864-3874.e6.

+ Open protocol

+ Expand

Isolation of Skin and Lymph Node Cells

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

Epidermal single cell suspensions were generated as described (35 ). Split dorsal and ventral sides of the ear pinna were floated on 2.5mg/ml Dispase II (Roche) in HBSS 2% FBS for 15 hours at 4°C, followed by mechanical dissociation of the epidermal layer in PBS/1mM EDTA/1% FBS by mincing with scissors or using the GentleMACS tissue dissociator (Miltneyi Biotech). Epidermal cells were passed sequentially through 70μM and 40μM cell strainers to remove clumps of cells. Numbers of cells were calculated by addition of counting beads (Invitrogen) before staining of samples for flow cytometry, and normalized to 0.1g weight for both complete ears before processing. After separation from the epidermis, the dermis was minced into small pieces and digested with 250 U/ml collagenase IV (Worthington, USA) and 800U/ml DNaseI (AppliChem, USA) at 37°C for 1 hour. Dermal single cell suspensions were then generated using the GentleMACS tissue dissociator (Miltneyi Biotech).
LN were teased apart with needles and digested in HBSS/4000U/ml collagenase IV (Worthington) for 40 min at 37°C. Digestion was quenched with 10mM EDTA, and the cells were passed through a 40 μM cell strainer, washed, and resuspended in PBS/1mM EDTA/1% FBS.
For blood cells, erythrocytes were removed by hypotonic lysis with distilled water, and cells then resuspended in PBS/1mM EDTA/1% FBS.

Ferrer I.R., West H.C., Henderson S., Ushakov D.S., Sousa P.S., Strid J., Chakraverty R., Yates A.J, & Bennett C.L. (2019). A wave of monocytes is recruited to replenish the long-term Langerhans cell network after immune injury.. Science immunology, 4(38), eaax8704.

+ Open protocol

+ Expand

Primary Sternal Chondrocyte Isolation

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

Primary sternal chondrocyte isolation was as described before.⁽⁴⁴⁾ In brief, mouse sterna and ribs from P3 neonates were cleaned and digested by pronase (Roche, Indianapolis, IN, USA; 11459643001) at 2 mg/mL PBS at 37°C with constant agitation for 1 hour, Collagenase IV (Worthington, Lakewood, NJ, USA; LS004189) at 3 mg/mL DMEM at 37°C for 1 hour, Collagenase IV at 0.5 mg/mL DMEM at 37°C for 3 hours, and filtered using 45 μm cell strainer.

Zhang H., Puviindran V., Nadesan P., Ding X., Shen L., Tang Y.J., Tsushima H., Yahara Y., Ban G.I., Zhang G., Karner C.M, & Alman B.A. (2022). Distinct Roles of Glutamine Metabolism in Benign and Malignant Cartilage Tumors With IDH Mutations. Journal of Bone and Mineral Research, 37(5), 983-996.

+ Open protocol

+ Expand

Isolation and Dissociation of Pancreatic Islets

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

Pancreata were perfused by injecting 3 mL collagenase 4 (Worthington, Lakewood, NJ) (400 units/mL in Hanks’ balanced salt solution (HBSS) and 10% fetal bovine serum (FBS)), harvested, and placed in 3–5 mL collagenase 4. The pancreata then were incubated at 37°C for 25 min, after which they were washed three times with 7 mL 5% FBS/HBSS and resuspended in 10 mL 5% FBS/HBSS. Islets were handpicked and incubated at 37°C for 15 min in 1 mL cell dissociation buffer (Invitrogen, Carlsbad, CA) and then further dissociated by vortexing and pipetting. Cells were then washed in 10 mL 5% FBS/HBSS, counted, and analyzed by flow cytometry.

Bettini M., Blanchfield L., Castellaw A., Zhang Q., Nakayama M., Smeltzer M.P., Zhang H., Hogquist K.A., Evavold B.D, & Vignali D.A. (2014). TCR affinity and tolerance mechanisms converge to shape T cell diabetogenic potential. Journal of immunology (Baltimore, Md. : 1950), 193(2), 571-579.

+ Open protocol

+ Expand

Phospho-Src Family Protein Analysis

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

Western blotting was performed as previously described. 12 The CTOSs in gel were digested by 2 mg/mL of collagenase IV (Worthington Biochemical Corporation) for 10 minutes, rinsed, and lyzed in the same way as the CTOSs in suspension. Primary antibodies against the phospho-Src family (Tyr416) and Src (36D10) were obtained from Cell Signaling Technology.

Okuyama H., Kondo J., Sato Y., Endo H., Nakajima A., Piulats J.M., Tomita Y., Fujiwara T., Itoh Y., Mizoguchi A., Ohue M, & Inoue M. (2016). Dynamic Change of Polarity in Primary Cultured Spheroids of Human Colorectal Adenocarcinoma and Its Role in Metastasis. The American journal of pathology, 186(4).

+ Open protocol

+ Expand

Isolation and Culture of Ovarian Cancer Cells

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

Human peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation, using Histopaque (Sigma-Aldrich, 10771), from buffy coats obtained from healthy blood donors (Blood Bank, University Hospital Basel, Switzerland). Fresh tumor samples were obtained from two ovarian cancer patients undergoing tumor resections at University Hospital Basel, Switzerland. Patient characteristics are summarized in Supplementary Table 1. The study was approved by the local Ethical Review Board (Ethikkommission Nordwestschweiz) and University Hospital Basel, Switzerland. Written consent to use their tumor samples for research purposes was obtained from all patients. Fresh tumor samples were mechanically dissociated and digested using accutase (Innovative Cell Technologies, AT-104), collagenase IV (Worthington, LS004188), hyaluronidase (Sigma-Aldrich, H6254), and DNAse type IV (Sigma-Aldrich, D5025), directly after excision. Single-cell suspensions were prepared and samples were stored in liquid nitrogen until further use. In the following assays, single-cell suspensions derived from ovarian cancer samples were maintained in RPMI medium containing L-glutamine (Sigma-Aldrich, R8758) supplemented with 1 × penicillin/streptomycin (Sigma-Aldrich, P4333) and 10% FBS (Pan Biotech, P30-5500).

Natoli M., Herzig P., Pishali Bejestani E., Buchi M., Ritschard R., Lloyd G.K., Mohanlal R., Tonra J.R., Huang L., Heinzelmann V., Trüb M., Zippelius A, & Kashyap A.S. (2021). Plinabulin, a Distinct Microtubule-Targeting Chemotherapy, Promotes M1-Like Macrophage Polarization and Anti-tumor Immunity. Frontiers in Oncology, 11, 644608.

+ Open protocol

+ Expand

Immune Cell Isolation from MC38 Tumors

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

For analysis of immune populations isolated from MC38 tumors, the following protocol was used. Harvested MC38 tumors were cut into smaller fragments with razor blades and then placed in 2–4 mL digestion mix [containing accutase (PAA), collagenase IV (Worthington), hyaluronidase (Sigma), and DNAse type IV (Sigma)] at 37°C for 45 min with constant shaking. Tumor suspensions were then filtered via 70 μM nylon mesh and washed with PBS containing EDTA (5 mM). Cell suspensions were subjected to red blood cell (RBC) lysis by adding 1 mL of RBC lysis buffer for 1 min, followed by a wash with PBS containing 2 mM EDTA. As a final step, cell suspensions were filtered via 70 μM nylon mesh and either stored in −80°C until further analysis or used directly for flow cytometry staining.

+ Open protocol

+ Expand

Isolation and Expansion of Marmoset Cell Lines

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

Animal care as well as all treatment procedures were in accordance with the current regulations as outlined in the animal protection law and reflected in the institutional guidelines. Marmoset cjFFs were isolated from leftover fragments of marmoset fetuses from days 70-74 of gestation, first used in other unrelated projects that have been fully approved by the Lower Saxony's State Office of Consumer Protection and Food Safety (LAVES) (license numbers 42502-04-16/2129 and 42502-04-16/2130) including a positive ethics evaluation. Tissues of the fetal dorsal body wall and limb buds were finely minced with scalpel blade and incubated in a mixture of 1:1 (v:v) Accutase (Gibco) and Collagenase IV (Worthington Biochemical Corporation)(2 mg/ml) at 37ºC for 15 min, followed by trituration to disaggregate the tissues to single cells or small clumps. The suspension was centrifuged at 300 g for 5 min, re-suspended in M20 culture medium (Dulbecco's DMEM containing GlutaMAX (Gibco) supplemented with 20% FBS (Gibco), non-essential amino acids (NEAA (Gibco), penicillin/streptomycin (Gibco), and 5 ng/ml bFGF), and plated on gelatin-coated 10 cm tissue culture dishes. The cells were subsequently split 1:4-1:6 with Accutase every 3-4 days and maintained in M20 medium.

Petkov S., Dressel R., Rodríguez-Polo I., & Behr R. (2020). Controlling the switch from neurogenesis to pluripotency during marmoset monkey somatic cell reprogramming with self-replicating mRNAs and small molecules.

+ 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!