Collagenase IV, CAS No.9001-12-1

CAS: 9001-12-1 Cat. No.: C1509223 Número CE: 232-582-9 PubChem CID: 44384985

GRADE & PURITY Bioactive, ActiBioPure™, Native, High Performance, EnzymoPure™, from Clostridium histolyticum; ≥125 U/mg enzyme powder

Accession #

Q46085,Q9X721

Bioactivity

≥125 U/mg enzyme powder

Visão geral

Collagenase is a protease derived from Clostridium histolyticum. It specifically recognizes the Pro-X-Gly-Pro sequence and cleaves the peptide bond between glycine (Gly) and glycine (Gly) in this sequence, and this sequence occurs frequently in collagen. Collagenase is the only protease that can degrade the triple-helical natural collagen fibers widely present in connective tissue.This product is Type IV collagenase, with an activity of ≥125 U/mg solid and low trypsin activity, making it particularly suitable for islet cell preparation.

Usage Instructions

Ⅰ. Preparation of Collagenase Stock Solution

Add 1 ml of the appropriate buffer (determined according to specific experimental conditions or relevant literature) to each tube containing 100 mg of collagenase (e.g., HBSS [Hank's Balanced Salt Solution, containing Ca²+, Mg²+, etc.]). Gently vortex to dissolve thoroughly and prepare a 100 mg/ml (i.e., 100×) stock solution. Filter-sterilize using a 0.22 μm filter with low protein binding properties, aliquot into small portions, and store frozen at -20°C in the dark.Thaw on ice before use; avoid repeated freeze-thaw cycles. The typical working concentration for tissue and cell dissociation is 0.5~2.5 mg/ml, and 1~2 mg/ml for cartilage digestion. Determine the optimal working concentration based on specific experimental conditions or relevant literature.

Ⅱ. Tissue Isolation

1. Use a sterile scalpel or scissors to cut the tissue into 3~4 mm tissue pieces;
2. Wash the tissue pieces several times with the appropriate buffer (e.g., HBSS);
3. Add sufficient buffer to submerge the tissue, then add collagenase to the required working concentration;
4. Incubate at 37°C for 4~18 hours. Using a horizontal shaker and supplementing digestion with 3 mM CaCl₂ can improve digestion efficiency;
5. Fully digested tissue can be filtered through a stainless steel or nylon mesh sieve and collected for later use. For incompletely dissociated tissue, add an appropriate amount of fresh collagenase working solution and continue incubation at 37°C;
6. Wash the collected cells several times with collagenase-free buffer;
7. Resuspend the cells in cell culture medium, and count the viable cell density using an automated cell counter or other methods;
8. Seed the cells on cell culture dishes using a suitable cell culture medium.

Ⅲ. Organ Perfusion

1. Add collagenase to the appropriate buffer (e.g., HBSS) preheated to 37°C; supplementing with 3 mM CaCl₂ helps improve isolation efficiency;
2. Perfuse the target organ with the collagenase working solution at an optimized rate;
3. Pass the recovered perfusion solution through a stainless steel or nylon mesh sieve to separate dissociated cells/small tissue fragments from larger clumps. Incompletely dissociated tissue should be further incubated with fresh collagenase working solution at 37°C;
4. Wash the collected cells several times with collagenase-free buffer;
5. Resuspend the cells in cell culture medium, and count the viable cell density using an automated cell counter or other methods;
6. Seed the cells on cell culture dishes using a suitable cell culture medium.

Specifications

Product Name

Collagenase IV, CAS No.9001-12-1

Sinónimos

microbial collagenase | clostridiopeptidase A | Clostridium histolyticum collagenase | collagenase A | collagenase Ⅳ | Collagenase type Ⅳ | Collagenase, Type Ⅳ | colG | colH

Grau

ActiBioPure™, Bioactive, High Performance, Native, EnzymoPure™

Especificações e pureza

Bioactive, ActiBioPure™, Native, High Performance, EnzymoPure™, from Clostridium histolyticum; ≥125 U/mg enzyme powder

Bioatividade

≥125 U/mg enzyme powder

Número de acesso

Q46085,Q9X721

CAS

9001-12-1

Número da Comissão das Enzimas

EC 3.4.24.3

Tipo de molécula

Enzyme

Product Line

Produtos biológicos

Armazenamento e envio

Concentração

from Clostridium histolyticum; ≥125 U/mg enzyme powder

Reconstituição

Reconstitute 100mg collagenase in 1 ml of of the corresponding buffer (determined according to specific experimental conditions or reference materials), e.g., HBSS (Hank's Balanced Salt Solution, containing Ca²+, Mg²+, etc.), and gently vortex to dissolve it thoroughly to prepare a 100 mg/ml (i.e., 100×) stock solution. Filter-sterilize using a 0.22 μm filter with low protein binding properties, aliquot into small portions, and store frozen at -20°C in the dark.

Condições de armazenamento de armazenamento

Store at 2-8°C

Enviado em

Wet ice

Estabilidade e armazenamento

Store at 2-8℃ long term (24 months). Upon reconstitution, it is recommended to aliquot.

Definição da unidade

One unit liberates one micromole of L-leucine equivalents from collagen in 5 hours at 37 ̊C, pH7.5.

Application Protocols

Item-specific

• No validated, item-specific protocols, dilutions, or compositions are provided here; refer to CoA/Spec Sheet.

Example workflows (general/literature; adapt empirically)

1. Mouse liver cell isolation (illustrative)

• Prepare collagenase working solution in HBSS + 10 mM HEPES + 5 mM CaCl2, chilled. Warm an aliquot to 37 °C immediately before use.
• Perfuse liver or mince tissue; incubate at 37 °C with gentle rocking for 10–20 min (e.g., 0.5–1.0 mg/mL). Monitor tissue softening.
• Quench with cold medium + 10% FBS; triturate gently; filter through 70 µm mesh; wash and proceed to density separation if needed.

2. Pancreatic islet enrichment (illustrative)

• Digest pancreas at 37 °C with collagenase solution (titrate within 0.5–1.5 mg/mL). Agitate gently 10–20 min.
• Quench and purify islets by gradient separation; minimize overdigestion to preserve viability.

3. Tumor dissociation for single-cell analysis (illustrative)

• Combine collagenase IV with DNase I and a neutral protease in HBSS/HEPES + Ca2+.
• Incubate minced tumor at 37 °C for 20–45 min with rotation. Intermittently pipet to assist dissociation.
• Quench, filter (40–70 µm), and proceed to RBC lysis and viability assessment.

General tips

• Always titrate enzyme concentration/time per tissue lot; maintain sterility; keep prep steps cold and digestion at target temperature; document lot-to-lot performance.

Biological Roles

General/literature context (not item-specific)

• Source and function: Bacterial collagenases (classically from Clostridium histolyticum) are zinc-dependent metalloproteases that cleave native triple-helical collagen, a major structural protein in animal extracellular matrices. In nature, such enzymes facilitate nutrient acquisition and tissue invasion by the producing microorganisms.
• Substrate specificity: Cleavage within Gly–X–Y repeats of collagens I–IV and others; once collagen is unwound or nicked, further degradation proceeds more readily (gelatinase-like activity may contribute).
• Ion dependence: Ca2+ stabilizes the enzyme tertiary structure and enhances catalytic performance; removal of divalent cations by chelators reduces activity.
• Biological impact in vitro: In controlled laboratory settings, collagenase digestion liberates cells embedded in collagen-rich matrices while often preserving more cell-surface markers than serine proteases.
• Interaction with ECM: Besides collagens, accessory proteolytic activities present in typical collagenase preparations can act on laminin, fibronectin, and proteoglycans to varying extents, aiding matrix loosening.
• Inhibition: Metal chelators (EDTA/EGTA), broad-spectrum metalloprotease inhibitors (e.g., 1,10-phenanthroline), and specific peptidic inhibitors can attenuate activity; endogenous tissue inhibitors of metalloproteinases (TIMPs) generally target MMPs rather than bacterial collagenases, but proteinaceous inhibitors and serum components can partially quench activity.

Note

• The above reflects typical biological properties of collagenase preparations; consult the CoA for the exact composition and accessory activities of this specific item.

Buffer Applications

Item-specific

• No buffer composition is specified for this item; refer to CoA/Spec Sheet. Storage at 2–8 °C per Product Data.

General/literature buffer practices (not item-specific)

• Common vehicles: HBSS (with Ca2+ and Mg2+), DPBS/PBS, or Krebs–Henseleit buffers. For CO2 incubations, bicarbonate-buffered media are suitable; for open-bench digestions, HEPES (10–25 mM) provides CO2-independent pH control.
• pH range: Maintain near neutral pH 7.2–7.6 during digestion to preserve activity and cell viability.
• Calcium supplementation: Include CaCl2 at 1–10 mM to support enzyme stability; avoid chelators (EDTA/EGTA) during active digestion.
• Stabilizers: 0.1–1 mg/mL BSA can reduce enzyme adsorption to plastics and stabilize activity; optional 0.5–1% serum may help quench nonspecific proteolysis post-dissociation.
• Sterility: Prepare solutions aseptically; filter through 0.22 µm membranes on ice. Pre-chill buffers to minimize premature activity before tissue addition.
• Quench/stop solutions: Cold media with serum (5–10%) or addition of EDTA (after desired digestion) can halt activity depending on downstream needs.

Recipe examples (literature; illustrative only)

• Collagenase working buffer: HBSS + 10 mM HEPES + 2–5 mM CaCl2 + 0.1% BSA. Adjust pH to 7.4, filter-sterilize cold.
• Quench medium: DMEM/F12 + 10% FBS + 10 mM HEPES, chilled on ice.

Green Alternatives

Context

• Collagenase IV is a biocatalyst enabling ECM degradation under mild, aqueous, near-neutral conditions—often replacing harsher chemical or thermal treatments.

Greenness assessment (literature)

• Advantages: Aqueous medium; ambient pressure; moderate temperatures (30–37 °C); reduced need for strong acids/bases or chaotropes; higher selectivity lowers waste from overdigestion.
• Tradeoffs: Enzyme manufacturing footprint; cold-chain logistics; potential for animal/bacterial source materials; activity loss requires careful handling, increasing consumables (ice/cooling).

Alternatives and comparisons (literature; not item-specific)

• Trypsin/EDTA: Widely available but less selective, more damaging to cell-surface proteins; EDTA conflicts with Ca2+-dependent processes and is not green for downstream if residual chelator is problematic.
• Dispase II or Neutral Protease: Gentler on surface epitopes but weaker against fibrillar collagens; may require longer digestions or combinations.
• Mechanical dissociation alone: No chemical inputs but often gives poor yields/viability for collagen-rich tissues; higher energy/processing time.
• Recombinant collagenases: Potential for more controlled impurity profiles and reduced animal-derived components; availability and cost vary.

Summary table (literature)

• Collagenase IV: High ECM selectivity; aqueous; moderate T; needs Ca2+; cold-chain required.
• Trypsin: Broad proteolysis; fast; can harm epitopes; EDTA often added (chelates metals).
• Dispase/Neutral protease: Gentle on cells; limited collagen cleavage; may need combo with collagenase.

Practical tip

• For greener workflows, optimize enzyme dose/time to minimize repeats, and implement reusable cold packs and insulated shipping to cut ice waste.

Pharmaceutical Uses

Item-specific

• For research use only (from Product Data). No clinical or diagnostic use is implied.

General/literature, non-clinical manufacturing context (not item-specific)

• Cell processing aid: Collagenase preparations are used in research-scale upstream processing to isolate primary cells (e.g., stromal vascular fraction from adipose, hepatocytes, islets) from collagen-rich tissues for in vitro research. In regulated manufacturing, enzymes must meet stringent specifications; those claims are not made here.
• Tissue engineering R&D: Employed to recover cells from 3D collagen or basement-membrane matrices and to passage organoids or spheroids embedded in hydrogel systems.
• Bioprocess sampling: Facilitates release of cells or proteins entrapped in collagenous scaffolds for analytical characterization.
• Excipients/status: No pharmacopeial status is claimed for this item. When pharmacopeial or GMP-grade enzymes are required, verify compliance and documentation separately.

Regulatory and quality notes

• For any process with potential translational intent, establish enzyme traceability, impurity profiles (endotoxin, host contaminants), and validated removal/inactivation steps post-digestion. This listing provides no such guarantees—consult CoA/Spec Sheet and your QA unit for suitability assessments.

Physical Properties

Item-specific (from Product Data)

• Physical constants (mp/bp, density, refractive index, logP): Not applicable to protein enzyme preparations; not specified for this item; refer to CoA/Spec Sheet.
• Appearance: Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties (not item-specific)

• Nature: Proteinaceous enzyme preparation; heterogeneous mixture by mass; not amenable to standard small-molecule physico-chemical constants.
• Solubility: Readily soluble in physiological aqueous buffers (e.g., PBS, HBSS) and salt solutions; insoluble in most organic solvents.
• pH activity profile: Broadly active in near-neutral ranges; typical activity optima reported near pH 7.0–7.6 (literature), with diminished activity under acidic conditions.
• Temperature profile: Active at physiological temperatures; typical use 30–37 °C. Thermal denaturation occurs on prolonged exposure to elevated temperatures; maintain on ice when preparing working solutions (literature guidance).
• Cofactors: Divalent cations, especially Ca2+, support stability/activity; chelators (EDTA/EGTA) inhibit activity (literature).
• Isoelectric and heterogeneity: Multiple protein species can present multiple pI values; exact values are preparation-dependent (literature).
• Activity units: Specific activity and definition of “Unit” vary by supplier and assay substrate; for this item, activity specifications are not provided here—refer to CoA/Spec Sheet.

Practical notes (literature)

• Avoid foaming and vigorous vortexing to reduce denaturation.
• Filter sterilization (0.22 µm) of chilled solutions is commonly feasible; prolonged filtration can reduce activity.

Quality and Grades

Item-specific (from Product Data)

• Grade/Designation included with this item: Bioactive; ActiBioPure™; Native; High Performance; EnzymoPure™.
• Research use: For research use only.

Interpretation of grades (general meanings; not item-specific specifications)

• Bioactive: Supplied to preserve native enzymatic activity under recommended storage and handling; minimal denaturation during manufacture.
• ActiBioPure™: Vendor-specific designation indicating enhanced purity/activity retention suitable for sensitive biological workflows.
• Native: Indicates non-recombinant, non-denatured enzyme preparation preserving native cofactors and post-translational features from the source material.
• High Performance: Emphasizes batch-to-batch consistency and robust performance in demanding applications (e.g., primary tissue dissociation) with controlled contaminant enzymes.
• EnzymoPure™: Vendor mark denoting an elevated level of enzyme purity and reduced endotoxin/contaminants relative to standard research grades (consult CoA for exact metrics).

What to verify on CoA/Spec Sheet (item-specific metrics not provided here)

• Specific activity definition and assay substrate
• Collagenase class composition (e.g., proportions of C. histolyticum collagenase I/II and accessory proteases)
• Protease/tryptic/neutral protease background
• Endotoxin bioburden (EU/mg) if relevant to cell isolation
• Bioburden/sterility status (filtered/sterile)
• Appearance, form (lyophilized vs solution), excipients, and recommended reconstitution buffer

Practical QC guidance (literature)

• Perform pilot digestions and titrations per tissue type; retain sample from each lot for comparability testing.

Reaction and Applications

Item-specific applications (from Product Data)

• Research Use Note: For research use only.

Expanded applications (general/literature; not item-specific claims)

• Primary tissue dissociation: Efficient digestion of extracellular matrix to liberate viable cells from tissues rich in basement membrane and interstitial collagen (e.g., liver, pancreas, adipose, lung, tumor, brain). Frequently paired with DNase I and a neutral protease (e.g., Dispase or Thermolysin) to balance yield and viability.
• Organoid/cell isolation: Facilitates release of epithelial fragments or organoids from Matrigel- or collagen-based matrices while maintaining higher epitope integrity than harsh serine proteases.
• ECM processing: Generates collagen peptides/gelatin for downstream biochemical assays (zymography substrates, mass spectrometry peptide mapping).
• Imaging and histology: Controlled partial digestion can improve antibody penetration in thick tissue sections for immunolabeling without over-damaging antigens.
• Bioprocess sample prep: Reduces viscosity and matrix entrapment during downstream processing of collagenous materials.

Practical considerations (literature)

• Titrate enzyme concentration (e.g., 0.05–2 mg/mL typical range) and incubation time (10–60 min) by tissue type; monitor cell viability and surface markers.
• Maintain temperature at 30–37 °C during digestion; prepare and keep working stocks on ice.
• Include CaCl2 (1–10 mM) unless contraindicated; avoid EDTA during active digestion.
• Quench by dilution, cooling, or addition of serum/defined inhibitor mixture after achieving desired dissociation.

Reaction Conditions

Item-specific

• No activity units, optimal pH, or formulation details are specified for this item; refer to CoA/Spec Sheet for lot-specific instructions.

General/literature conditions for tissue dissociation (not item-specific)

• Concentration: Common working ranges 0.05–2.0 mg/mL, tuned by tissue density and desired gentleness.
• Temperature: 30–37 °C for active digestion; prepare and hold solutions on ice prior to use.
• pH/Buffer: Near-neutral pH 7.2–7.6 in HBSS/DPBS/PBS or HEPES-buffered saline.
• Cofactors: Include CaCl2 at 1–10 mM to support activity; Mg2+ may be present as part of balanced salts.
• Additives: DNase I (5–100 µg/mL) to reduce viscosity; neutral protease/Dispase to broaden ECM cleavage while preserving epitopes.
• Time: 10–60 minutes typical; longer incubations may increase yield at the expense of viability/epitopes.
• Agitation: Gentle rocking or slow rotation improves mass transfer; avoid vigorous shear that damages cells.
• Quenching: Dilution with cold serum-containing medium, cooling on ice, and/or addition of chelators (post-digestion) to halt activity.

Quality controls (literature)

• Monitor single-cell yield, viability (e.g., Trypan Blue), and marker retention by flow cytometry.
• Pilot titrations across concentrations/timepoints are recommended per tissue and lot to standardize outcomes.

Safety and Handling

Item-specific (from Product Data)

• GHS Classification, Signal Word, H-statements, Pictograms: Not specified for this item; refer to the product SDS for authoritative safety data.
• Storage: 2–8 °C. Shipped on wet ice (do not freeze unless CoA/SDS indicates suitability).

General/literature safety guidance (not item-specific)

• Hazard overview: Protein enzymes may cause respiratory or skin sensitization upon repeated exposure; avoid aerosol generation and inhalation of powders or droplets. Collagenase solutions can irritate eyes/skin.
• PPE: Lab coat, gloves (nitrile), and safety glasses. Use a certified biosafety cabinet for solution prep and tissue dissociation steps that can generate aerosols.
• Handling: Work on ice for solution prep to preserve activity. Open lyophilized vials carefully to prevent airborne dust. Avoid contact with oxidants and protease inhibitors (e.g., PMSF, EDTA) unless intentionally quenching.
• Incompatibilities: Chelators (EDTA/EGTA), heavy-metal chelating buffers, and strong oxidants degrade/inhibit activity. Avoid protease inhibitor cocktails during digestion phases.
• First aid (summary; defer to SDS): Inhalation—move to fresh air; seek medical attention if symptoms persist. Skin—wash with soap/water. Eyes—rinse with water for 15 minutes; remove contact lenses. Ingestion—not intended for human consumption; rinse mouth and seek medical advice.
• Spill response: Avoid raising dust/aerosols; damp wipe and disinfect surfaces. Dispose according to institutional biosafety procedures.
• Waste: Treat as bioactive protein waste; deactivate enzymatic activity (e.g., by adding EDTA and heat) prior to disposal if permitted by local regulations.

Solvent Selection

Item-specific

• This enzyme preparation is intended for aqueous use. Organic solvent systems are not applicable.

General/literature guidance (not item-specific)

• Primary media: Physiological salt solutions (e.g., HBSS, DPBS, PBS) with or without Mg2+/Ca2+ depending on protocol. Ca2+ (1–10 mM) is commonly included to support activity and stability.
• Buffering: HEPES (10–25 mM) or bicarbonate/CO2 systems help maintain near-neutral pH (7.2–7.6). Avoid phosphate concentrations that precipitate with Ca2+ at high levels.
• Incompatibilities: Chelating agents (EDTA/EGTA), high citrate, or phosphate at combinations that reduce free Ca2+ will inhibit enzyme activity. Detergents and strong denaturants (SDS, urea) inactivate proteins.
• Additives: BSA (0.1–1 mg/mL) can reduce nonspecific adsorption and stabilize activity. DNase I is frequently co-formulated to reduce viscosity from released DNA during tissue dissociation.
• Sterility: Filter-sterilize chilled solutions through 0.22 µm PVDF/CA membranes; protein-binding can be mitigated with BSA.

Comparison snapshot (literature)

• Collagenase IV vs Dispase II: Collagenase IV provides broader collagen cleavage for dense ECM; Dispase retains cell–cell junctions better but is weaker on fibrillar collagens.
• Collagenase IV vs Trypsin: Collagenase targets ECM collagens, preserving many surface epitopes; trypsin is a broad serine protease more likely to cleave cell-surface proteins.

Storage and Reconstitution

Item-specific (from Product Data)

• Storage conditions: Store at 2–8 °C.
• Shipping: Shipped on wet ice.
• Formulation/appearance/reconstitution instructions: Not specified for this item; refer to CoA/Spec Sheet.

General/literature guidance (not item-specific)

• Upon receipt: Keep refrigerated (2–8 °C). If lyophilized, allow vial to equilibrate to room temperature in a desiccated environment before opening to prevent condensation.
• Reconstitution: Prepare on ice using sterile buffer (e.g., HBSS or PBS/HEPES) supplemented with CaCl2 (1–10 mM). Optional 0.1% BSA can stabilize the enzyme and reduce adsorption.
• Concentrated stocks: Make a chilled, sterile stock solution; filter through 0.22 µm if sterility is required and compatible with your workflow.
• Aliquoting and storage: Divide into single-use aliquots to avoid repeated freeze–thaw. Short-term storage of working solutions at 2–8 °C for the day of use; for longer-term storage, follow CoA/SDS. Many labs freeze at −20 °C or −80 °C with stabilizers, but stability is preparation-dependent.
• Handling: Avoid vigorous mixing/foaming. Use low-protein-binding plastics when possible.
• Inactivation/cleanup: After use, quench activity by cooling/dilution and/or adding serum or EDTA (post-digestion) as appropriate. Clean spills with protein denaturants and standard disinfectants.

Always defer to the product-specific CoA and SDS for definitive storage, stability, and reconstitution instructions.

Structure and Identity

Item-specific (from Product Data)

• Product name: Collagenase IV (SKU C1509223)
• CAS: 9001-12-1
• Grade/Designation: Bioactive; ActiBioPure™; Native; High Performance; EnzymoPure™
• Storage: 2–8 °C; shipped on wet ice
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

General/literature description (not item-specific)

• Identity/Type: Collagenase IV typically refers to a preparation of collagen-degrading proteases commonly sourced from Clostridium histolyticum, enriched for activities that efficiently digest basement membrane and interstitial collagens alongside accessory proteases.
• Macromolecular nature: Protein enzyme(s), not a single small-molecule structure; thus SMILES/InChIKey are not applicable in the conventional sense.
• Structural features: Proteins with catalytic zinc-binding metalloprotease domains (HELGH motif variants) in the collagenase components; activity is generally Ca2+-dependent for stability and optimal conformation.
• Substrate scope: Triple-helical collagens (e.g., types I–IV) and gelatin; accessory proteases may broaden specificity to non-collagen ECM proteins.
• Form: Supplied as a protein preparation (powder or solution, depending on lot/vendor). Exact appearance for this item is not specified; refer to CoA/Spec Sheet.

2D/functional depiction in words (literature)

• Enzymatic cleavage of collagen’s Gly–X–Y repeats within the triple helix, followed by gelatinase-like secondary breakdown of denatured fragments.

Synthetic Utility

Applicability

• As a protein enzyme targeting biological collagen, Collagenase IV is not a typical reagent for small-molecule synthetic chemistry. Classical synthetic transformations (e.g., cross-coupling, reductions) do not apply.

Relevant laboratory utility (general/literature)

• Biocatalysis of biomaterials: Enables controlled depolymerization of collagen-based materials to peptides/gelatin, which can be analyzed by LC–MS or used to generate defined ECM fragments for biochemical assays.
• Sample preparation: Facilitates release of analytes or cells from collagenous matrices prior to omics workflows (proteomics, metabolomics) where minimal chemical modification is desired.
• Proteomics mapping: Limited, controlled digestion of collagen can generate fragments for structural studies distinct from tryptic peptides, aiding sequence coverage of collagen’s heavily modified domains.

Guidance

• If the goal is polymer/biomaterial modification, collagenase provides a biologically selective route compared to acid/base hydrolysis. For small-molecule synthesis, choose conventional organic reagents rather than this enzyme.

Target Specificity

Item-specific

• This product is not an antibody; no antigen/epitope/isotype data apply. Target specificity metrics are not specified for this item; refer to CoA/Spec Sheet.

General/literature substrate specificity for collagenase preparations (not item-specific)

• Primary substrates: Native triple-helical collagens, notably types I, II, III, and IV. Collagenase IV preparations are often selected for efficient basement membrane and interstitial collagen digestion.
• Cleavage characteristics: Endopeptidic scission within Gly–X–Y repeats; after initial nicking, gelatin-like fragments are further degraded. Accessory proteases may extend activity to laminin, fibronectin, and proteoglycans, facilitating matrix loosening.
• Inhibition profile: Activity reduced by chelation of divalent cations (EDTA/EGTA) and metal-binding inhibitors; resistant to classic serine protease inhibitors that target trypsin-like enzymes.
• Practical implication: Collagenase IV often preserves more cell-surface epitopes than broad serine proteases during cell isolation, improving downstream immunostaining/flow performance (protocol-dependent).