Collagenase dispase enzyme mix

Manufactured by Roche

Collagenase/dispase enzyme mix is a laboratory reagent used for the dissociation and isolation of cells from tissues. It is a combination of two enzymes, collagenase and dispase, that work together to break down the extracellular matrix, allowing for the effective release of cells from their surrounding environment.

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Lab products found in correlation

Blasticidin, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium, by Corning (1 mentions)

Spelling variants of this product

Collagenase (359 citations) Dispase (328 citations) Collagenase/dispase (301 citations)

2 protocols using collagenase dispase enzyme mix

Isolation and Culturing of Obsl1-Knockout Cells

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Culture of cos-1 cells was done as previously described^{76 (link),77 (link)}. To test Obsl1 antibodies and to determine if effects seen in dKO and Obsl1-knockout muscles are also seen in non-muscle Obsl1-knockout cells, we isolated cells from lungs of heterozygous global Obsl1 knockouts (+/−) and from mice that have a knockout and a floxed allele for Obsl1 (flox/−), using a collagenase/dispase enzyme mix (Roche). Isolated cells were cultured and propagated in growth medium (10% fetal calf serum, 1% penicillin/streptomycin, Dulbecco’s modified Eagle’s medium [Corning]). To generate Obsl1 knockouts, cells were transduced with a lentivirus carrying a cre-blasticidin-RFP expression cassette under control of a CMV promoter (#LVP013; Gentarget Inc.), allowing for recombination of the Obsl1-floxed allele and selection of transduced cells by 5 µg/ml blasticidin (Thermo Fisher). After transduction, cells were kept in growth medium supplemented with blasticidin until harvest for analysis of proteins.

Blondelle J., Marrocco V., Clark M., Desmond P., Myers S., Nguyen J., Wright M., Bremner S., Pierantozzi E., Ward S., Estève E., Sorrentino V., Ghassemian M, & Lange S. (2019). Murine obscurin and Obsl1 have functionally redundant roles in sarcolemmal integrity, sarcoplasmic reticulum organization, and muscle metabolism. Communications Biology, 2, 178.

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Isolation and FACS sorting of HER2-positive cells

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Tumor tissue was minced in RPMI and incubated in RPMI + 10% FBS + 1% penicillin/streptomycin (P/S) with collagenase/dispase enzyme mix (1 mg/ml; Roche) at 37°C, shaking for 1 hour. Tissue was then centrifuged at 1500 rpm for 5 min, and supernatant was removed. Pellet was resuspended in RPMI + 10% FBS + 1% P/S and pipetted well to dissociate clumps. Medium was refreshed the following day. FACS was performed using FACSAria III Fusion SORP. HER2-positive cells were sorted with a phycoerythrin-conjugated HER2 antibody (eBiosciences).

Kodack D.P., Askoxylakis V., Ferraro G.B., Sheng Q., Badeaux M., Goel S., Qi X., Shankaraiah R., Cao Z.A., Ramjiawan R.R., Bezwada D., Patel B., Song Y., Costa C., Naxerova K., Wong C.S., Kloepper J., Das R., Tam A., Tanboon J., Duda D.G., Miller C.R., Siegel M.B., Anders C.K., Sanders M., Estrada M.V., Schlegel R., Arteaga C.L., Brachtel E., Huang A., Fukumura D., Engelman J.A, & Jain R.K. (2017). The brain microenvironment mediates resistance in luminal breast cancer to PI3K inhibition through HER3 activation. Science translational medicine, 9(391), eaal4682.

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