Buyer guide

Collagenase Type II: Powder or Ready Solution - Activity Units, Working Concentration and Digestion Time

A tissue dissociation that gives too few cells, or dead ones, wastes the tissue, the animals and a working day, and the enzyme lot is often the reason. Collagenase type II is sold here in two formats: a 100 mg vial of powder kept at 2 to 8 °C, and a frozen 10 mL bottle of ready solution at 1 mg/mL. This page is for distributors, resellers and procurement buyers: which format fits which customer lab, what the activity number on a quote does and does not tell you, and what to ask us for before you request a wholesale quote.

Buying for resale, private label or bulk supply? Email [email protected] or use the quote form to request a wholesale quote — our team replies within one business day with pack options and wholesale pricing.

Powder or ready solution: the purchasing facts

	Type II powder	Type II ready solution
Pack	100 mg vial	10 mL bottle at 1 mg/mL
Storage	2 to 8 °C, protected from light	-30 to -5 °C, protected from light
Shipping class	Blue ice	Dry ice
Shelf life	24 months	24 months
Raw materials	Animal origin-free	Animal origin-free
Endotoxin	Not listed; ask for the lot figure	Not more than 10 EU/mL
Work on your side	Weigh, dissolve, filter, aliquot	Thaw and use

Enzyme per pack. A 10 mL bottle at 1 mg/mL holds a tenth of the enzyme in a 100 mg vial, so one vial of powder equals ten bottles of solution.
Cost per milligram. At list price the powder costs about one third as much per milligram as the ready solution; the solution is the convenience format and the powder is the volume format.
Freight. The powder ships on blue ice and the solution on dry ice. Dry ice costs more to ship and is more restricted. Freight is quoted per order, so ask for both options if you are undecided.
Storage space. The powder needs fridge space at 2 to 8 °C; the solution needs freezer space at -30 to -5 °C.
Shelf life. Both carry 24 months as supplied, so shelf life does not decide the format.
Who should buy which. A lab that digests tissue every week and has a balance, a filter and someone to aliquot buys powder. A lab that runs an occasional isolation, or wants the same ready bottle on every bench, buys the 1 mg/mL solution.

The activity units trap

Buyers compare collagenase quotes by units per milligram, and that is the comparison most likely to mislead. A specialist webinar on these enzymes makes the point directly: the long-used fibre-digestion assay dates from the 1950s and, in the presenters' words, "has never been characterized", with no published linearity, precision or specificity. A kinetic fluorescent assay for collagen degradation activity was developed in 2008, and enzyme mixtures with higher specific activity in that assay gave higher islet yields. The same webinar reports that intact class I collagenase shows 8-10 fold higher specific collagen degradation activity than intact class II or truncated class I. It also states: "Class I and class II collagenase are needed for the synergistic degradation of collagen."

A units figure is only comparable with another figure from the same assay, so ask every supplier for the assay name with the number.
A crude collagenase "type" is a blend, and the balance of the two classes changes how a lot behaves on your tissue.
Our listings for the powders and solutions state no activity figure, so we do not print one here. Ask us for the lot data and we tell you what is available for that lot.

Working concentration and digestion time

None of our collagenase listings states a working concentration or a digestion time, and we will not guess one for your tissue. Both depend on the tissue, the species, the age of the animal and the activity of the lot, so the starting point should come with the lot figure, and a new lot should be titrated on a small piece of tissue before a full run. The only fixed number is the format: the ready solution is supplied at 1 mg/mL, so any dilution starts from there. Tell us the tissue and the cell number you need and we will suggest the type; ask us for the lot data and we tell you what is available for that lot.

A protocol does not transfer between tissues without re-testing, and the enzyme order should follow the tissue.

Which type, and what goes in the bottle with it

We list three collagenase types, each as a 100 mg powder and as a 10 mL ready solution at 1 mg/mL, with the same pack and price pattern. Our dispase and collagenase reference gives the usual match: type I for general work on soft tissue, type II for heart, bone, muscle and cartilage, type IV for islets and other cells that need low tryptic activity. Islet isolation is the classic demanding case; a published human islet protocol calls quality and quantity of islets a prerequisite for transplantation and uses an automated method.

Collagenase needs calcium, and EDTA takes it away. That matters because the trypsin products on the same order contain EDTA. Do not carry trypsin-EDTA or a calcium-free rinse into a collagenase step.

Reagent	Pack	Listed additions	Storage	Shipping	Shelf life
Collagenase type I powder	100 mg	None	2 to 8 °C	Blue ice	24 months
Collagenase type II powder	100 mg	None	2 to 8 °C	Blue ice	24 months
Collagenase type IV powder	100 mg	None	2 to 8 °C	Blue ice	24 months
Collagenase type I solution, 1 mg/mL	10 mL	None	-30 to -5 °C	Dry ice	24 months
Collagenase type II solution, 1 mg/mL	10 mL	None	-30 to -5 °C	Dry ice	24 months
Collagenase type IV solution, 1 mg/mL	10 mL	None	-30 to -5 °C	Dry ice	24 months
Trypsin-EDTA 0.25%	500 mL	2.5 g/L trypsin, 0.38 g/L EDTA	-30 to -5 °C	Dry ice	24 months
Trypsin-EDTA 0.25%, no phenol red	100 mL	Not listed	-30 to -5 °C	Dry ice	24 months
Trypsin-EDTA 0.05%	500 mL	0.5 g/L trypsin, 0.2 g/L EDTA	-30 to -5 °C	Dry ice	24 months
Recombinant trypsin 10X	500 mL	1 mM EDTA	2 to 8 °C	Blue ice	36 months
Trypsin 1:250 powder	100 g	None	2 to 8 °C	Blue ice	24 months

The pattern for the order: powders travel cold on blue ice, ready enzyme solutions travel frozen on dry ice, so a mixed order ships in two temperature classes.

Loose enzyme, ready kit or instrument

A buyer today is offered three routes to a single-cell suspension. Ready tissue dissociation kits are promoted as effortless enzymatic dissociation. Benchtop dissociators are sold on ease. Dedicated single-cell suspension instruments and semi-automated disaggregators for solid tissue and tumour samples are aimed at high-throughput work, single-cell sequencing and flow cytometry. Kits and instruments buy speed and repeatability and tie the lab to one consumable. Loose enzyme is the lowest cost per digestion and the most flexible, and a ready 1 mg/mL solution sits between the two: no weighing, no hardware.

Before you digest: a short checklist

Name the tissue and the target cell. The type follows the tissue.
Ask for the lot data and the assay name. We tell you what is available for that lot; without the assay name a units figure cannot be compared.
Decide the format. Powder for regular use, the 1 mg/mL solution for occasional runs.
Check the diluent. Use a buffer or medium with calcium, and keep EDTA out.
Titrate the new lot. Test on a small piece of tissue before the full run.
Plan the cold chain. Fridge for powder, freezer for solution, and dry ice delivery for the solution.

Mistakes that cost tissue and money

Comparing units per milligram across suppliers. Different assays give numbers that do not compare, and the oldest one was never characterised.
Assuming a new lot behaves like the last one. The class balance matters, with 8-10 fold differences in specific activity between forms of the enzyme.
Using the wrong type for the tissue. Three types are listed because tissues differ.
Carrying EDTA into the digestion. The trypsin-EDTA products list EDTA in the formula; keep them apart from the collagenase step.
Copying a protocol from another tissue. A method for one tissue, region, animal age or species needs re-testing on the next.
Ordering solution with no freezer space or dry ice receiving. The solution is stored at -30 to -5 °C and ships on dry ice.
Buying ten bottles when one vial would do. One 100 mg vial holds the enzyme of ten 10 mL bottles at 1 mg/mL.

Request a wholesale quote, the spec sheet and the lot data

Email [email protected] to request a wholesale quote or the spec sheet for collagenase type I, II or IV as powder or solution, or to ask about a trial quantity. Ask us for the lot data and we tell you what is available for that lot. Tell us:

the tissue, species and target cell;
the cell number you need per run and runs per month;
powder or solution, and the delivery country;
whether you buy for resale, private label or your own use.

We reply with a suggested type and wholesale pricing.

Reference pages on this topic

DispaseDispase, also sold as Dispase II or neutral protease, is a zinc-dependent metalloprotease from Paenibacillus (formerly Bacillus) polymyxa, EC 3.4.24.4, used for gentle tissue dissociation and for detaching cells and epithelial sheets intact. It cleaves fibronectin and type IV collagen, degrades type I collagen only minimally, and does not cleave laminin or type V collagen, which is why it can separate an epidermis from a dermis or lift a confluent epithelial sheet off plastic without destroying cell-cell junctions. Because it is a metalloprotease requiring zinc for catalysis and calcium for stability, it is inhibited by EDTA, EGTA and 1,10-phenanthroline, and it must be diluted in a calcium-containing buffer rather than a chelator-based one.
Puromycin for Mammalian Cell SelectionPuromycin is an aminonucleoside antibiotic from Streptomyces alboniger that mimics the aminoacyl end of tRNA, enters the ribosomal A site and causes premature release of a truncated peptide chain, killing cells that cannot inactivate it. In mammalian cell culture it is used as a selection agent at 0.5-10 ug/mL, most commonly 1-2 ug/mL, and it kills non-resistant cells faster than any other common selection antibiotic -- typically within 2-5 days. Resistance is conferred by the pac gene encoding puromycin N-acetyltransferase, which acetylates the drug and renders it inactive. Because the effective concentration depends strongly on cell line, cell density and serum content, a kill curve must be run for each new cell line and each new lot of puromycin.
Penicillin-Streptomycin and Antibiotic-Antimycotic in Cell CulturePenicillin-streptomycin, universally shortened to pen-strep, is supplied as a 100X sterile solution containing 10,000 units/mL penicillin G and 10,000 ug/mL streptomycin, diluted 1:100 into medium to give a working concentration of 100 U/mL penicillin and 100 ug/mL streptomycin. Penicillin blocks bacterial cell wall synthesis and covers mainly Gram-positive organisms; streptomycin binds the bacterial 30S ribosomal subunit and covers mainly Gram-negatives, so the pair gives broad antibacterial coverage. Antibiotic-antimycotic (anti-anti) is the same combination plus 25 ug/mL amphotericin B at 100X, giving 0.25 ug/mL in use for antifungal coverage. Neither controls mycoplasma, and major cell banks including ATCC recommend against routine antibiotic use because it masks low-level contamination rather than preventing it.
Antibiotics in Cell Culture: Working Concentrations and When Not to Use ThemAntibiotics in cell culture are antibacterial and antifungal agents added to growth medium to suppress microbial contamination; the standard combinations are penicillin–streptomycin at 100 U/mL and 100 µg/mL, gentamicin at 5–50 µg/mL, amphotericin B as an antimycotic at 0.25–2.5 µg/mL, and antibiotic–antimycotic 100X, which supplies all three at 100 U/mL, 100 µg/mL and 0.25 µg/mL when diluted. The professional consensus is that these should not be used routinely or prophylactically: they mask low-level contamination, select for resistant organisms, do nothing at all against mycoplasma, and can alter cell physiology and experimental response. Their defensible uses are primary tissue isolation, work with material from non-sterile sites, and short-term rescue of an irreplaceable culture — not the standard maintenance of established cell lines.
RPMI 1640 MediumRPMI 1640 is a basal cell culture medium developed at Roswell Park Memorial Institute in 1966 for the culture of human leukocytes in suspension. It contains 2,000 mg/L glucose (11.1 mM), 2,000 mg/L sodium bicarbonate (23.8 mM) buffered for a 5% CO2 atmosphere, unusually high phosphate (about 5.6 mM), low calcium (about 0.42 mM), and a distinctive component set that includes reduced glutathione, biotin, vitamin B12, para-aminobenzoic acid and hydroxyproline. It is the standard medium for lymphocytes, hybridomas, and most suspension-adapted haematopoietic and lymphoid cell lines, normally supplemented with 10% fetal bovine serum.
DMEM (Dulbecco's Modified Eagle Medium)DMEM (Dulbecco's Modified Eagle Medium) is a basal cell culture medium derived from Eagle's Minimal Essential Medium by raising the amino acid and vitamin concentrations roughly fourfold. It is supplied in high-glucose (4,500 mg/L, 25 mM) and low-glucose (1,000 mg/L, 5.6 mM) forms, buffered with 3,700 mg/L sodium bicarbonate, and requires serum or a defined supplement plus a CO2 atmosphere to hold physiological pH. DMEM is the default medium for adherent lines such as HEK293, HeLa, NIH/3T3, Vero and CHO-derived adherent cultures, and for most primary fibroblasts.

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