Implantation sites were harvested as described above. Each site was cut into 12 pieces and placed in RPMI 5% fetal bovine serum (FBS). These pieces were digested in RPMI containing 5% FBS, 300 μg/ml collagenase F (Sigma-Aldrich), 200 μg/ml collagenase L (Sigma-Aldrich), 500 μg/ml Dispase (Gibco), and 2 U/ml DNase-1 (Roche) at 37°C for 30 to 45 min with a magnetic stir bar for agitation. Cells were passed over a 70 μm strainer (BD) to create a single cell suspension. Implantation sites were washed in DPBS, 1% FBS, 25 mM EDTA. Cells were stained for FACS with anti-CD45 (30-F11, BD), anti-CD11b (M1/70, BD), anti-Gr-1 (RB6-8C5, BD), and rabbit anti-Crry followed by a donkey anti-rabbit DyLight 488 (Jackson ImmunoResearch). Blocking for FACS was carried out employing DPBS, 1% FBS, 25 mM EDTA with 5% donkey serum and 5% mouse serum. Cells were examined employing a FACScan (BD) retrofitted with a Cytek Upgrade.
Collagenase l
Manufactured by Merck Group
Collagenase L is a laboratory enzyme used for the digestion of collagen, a structural protein found in various tissues. It is a complex of enzymes that work together to break down collagen into smaller peptides. The core function of Collagenase L is to facilitate the isolation and extraction of cells from collagen-rich environments, such as connective tissues, for further biological research and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Dnase 1, by Roche (2 mentions)
Anti cd11b m1 70, by BD (2 mentions)
Anti gr 1 rb6 8c5, by BD (2 mentions)
Collagenase f, by Merck Group (2 mentions)
Facscan, by BD (2 mentions)
70 μm strainer, by BD (2 mentions)
Dispase, by Thermo Fisher Scientific (2 mentions)
Anti cd45 30 f11, by BD (2 mentions)
Dylight 488, by Jackson ImmunoResearch (2 mentions)
2 protocols using collagenase l
1
Isolation and Analysis of Implantation Site Cells
2
Isolation and Analysis of Implantation Site Cells
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Implantation sites were harvested as described above. Each site was cut into 12 pieces and placed in RPMI 5% fetal bovine serum (FBS). These pieces were digested in RPMI containing 5% FBS, 300 μg/mL collagenase F (Sigma‐Aldrich), 200 μg/mL collagenase L (Sigma‐Aldrich), 500 μg/mL Dispase (Gibco), and 2 U/mL DNase‐1 (Roche) at 37°C for 30‐45 minutes with a magnetic stir bar for agitation. Cells were passed over a 70 μm strainer (BD) to create a single‐cell suspension. Implantation sites were washed in DPBS, 1% FBS, 25 mmol/L EDTA. Cells were stained for FACS with anti‐CD45 (30‐F11, BD), anti‐CD11b (M1/70, BD), anti‐Gr‐1 (RB6‐8C5, BD), and rabbit anti‐Crry followed by a donkey anti‐rabbit DyLight 488 (Jackson ImmunoResearch). Blocking for FACS was carried out employing DPBS, 1% FBS, 25 mmol/L EDTA with 5% donkey serum, and 5% mouse serum. Cells were examined employing a FACScan (BD) retrofitted with a Cytek Upgrade.
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