Bronchoalveolar lavage (BAL) was performed by cannulating the trachea, injecting 1 mL of PBS and subsequently aspirating the BAL fluid. Liberase/DNAse I (both Sigma-Aldrich) digests of the lung were prepared to obtain single lung cell suspensions (21 (link)). Single lung cell suspensions were used for the analysis of phagocyte subpopulations by flow cytometry.
Liberase dnase 1
Manufactured by Merck Group
Liberase/DNAse I is a laboratory reagent used for the isolation and dissociation of cells from tissue samples. It contains a proprietary blend of enzymes, including collagenase and deoxyribonuclease I, which work together to break down extracellular matrix components and DNA, facilitating the release of individual cells from the tissue. This product is commonly used in cell biology research, tissue engineering, and regenerative medicine applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti human cd45 apc, by Miltenyi Biotec (1 mentions)
Anti human cd45 microbeads, by Miltenyi Biotec (1 mentions)
Hla dr pe, by Miltenyi Biotec (1 mentions)
Facscanto 2 cell analyzer, by BD (1 mentions)
Foxp3 staining buffer set, by Thermo Fisher Scientific (1 mentions)
Automacs pro separator, by Miltenyi Biotec (1 mentions)
2 protocols using liberase dnase 1
1
Isolation of Lung Cell Subsets
2
Isolation and Characterization of Human Thymic Cells
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Human thymus samples were collected from pediatric patients undergoing cardiac surgery (under the clinical protocol TIGET07, approved by the Ethical Committee of San Raffaele Scientific Institute). Thymic tissue was cleaned and minced into small pieces before digestion with Liberase/DNase I (Sigma-Aldrich). Digested tissue was depleted of CD45-positive cells using the AutoMACS Pro separator (Miltenyi Biotec) after incubation with anti–human CD45 microbeads (Miltenyi Biotec). After depletion, thymic cells were stained with antibodies anti–human CD45 APC, epithelial cell adhesion molecule/CD326 Pe-Vio770, HLADR PE, CD31 APC-Vio770 (all from Miltenyi Biotec), and Ulex europaeus agglutinin I FITC (UEA I FITC; Vector Laboratories). After surface staining, thymic cells were fixed/permeabilized (Foxp3 Staining Buffer set; eBioscience) and stained intracellularly with anti-Extl3 antibody (Ab113063; Abcam), followed by a secondary goat anti–rabbit IgG Alexa Fluor 405 antibody (Invitrogen). Then, cells were acquired on a FACS Canto II cell analyzer (BD) and analyzed using FlowJo software (Tree Star).
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