For flow cytometry analysis, cells were washed and incubated with Fc-receptor blockage using CD16/CD32 (Mouse BD Fc Block) (clone 2.4G2 (RUO), BD Pharmingen) and 5% rat serum (Invitrogen, Carlsbad, CA, USA) for 10 min prior to labeling. Subsequently, the cells were incubated with labelling antibodies for 20 min, washed and analyzed on the Attune NxT Acoustic Focusing Cytometer (Thermo Fisher Scientific). The following fluorescent-labeled antibodies were used for cell labeling: Ly6G (clone: 1A-8), ICAM-1 (CD54) (clone: 3E2), CD62L (L-Selectin) (clone: MEL-14), CXCR1 (clone: U45-632), all from BD Biosciences and CD11b (clone: M1/70) and CXCR2 (clone: SA044-G4), from BioLegend (San Diego, CA, USA). Neutrophil subsets were characterized using CD62L (L-Selectin) and ICAM-1 (CD54). Neutrophils were considered immature when expressing a CD62L + /ICAM-1(CD54)-phenotype and mature when expressing CD62L + /ICAM-1(CD54) + or CD62L-/ICAM-1(CD54) + 21 (link),22 (link).
Icam 1 cd54
Manufactured by BD
Sourced in United States, France
ICAM-1 (CD54) is a cell adhesion molecule that mediates intercellular adhesion. It is expressed on the surface of endothelial cells, immune cells, and other cell types. ICAM-1 plays a role in leukocyte recruitment and cellular interactions during immune responses and inflammatory processes.
Automatically generated - may contain errors
Lab products found in correlation
Rat serum, by Thermo Fisher Scientific (2 mentions)
Attune nxt acoustic focusing cytometer, by Thermo Fisher Scientific (2 mentions)
Fluorochrome conjugated mouse igg isotype control, by BD (2 mentions)
Endoglin cd105, by BD (2 mentions)
Guava easycyte ht flow cytometer, by Merck Group (2 mentions)
Pecam1 cd31, by BD (2 mentions)
Cxcr1, by BD (1 mentions)
Mouse fc block, by BD (1 mentions)
Cxcr2, by BioLegend (1 mentions)
Cd11b, by BioLegend (1 mentions)
Mycostrip mycoplasma detection kit, by InvivoGen (1 mentions)
L selectin cd62l, by BD (1 mentions)
Act 10, by Beckman Coulter (1 mentions)
Sigmafast bcip nbt, by Merck Group (1 mentions)
Stem cell kit, by STEMCELL (1 mentions)
Alcian blue dye, by Merck Group (1 mentions)
6 protocols using icam 1 cd54
1
Neutrophil Subtyping by Flow Cytometry
2
Endothelial Cell Surface Phenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The surface phenotype of EA.hy926 endothelial cells was determined by flow cytometry. Cells were fixed as described above. Direct labelling was performed for the following markers: endoglin/CD105 (BD Biosciences, Le Pont de Claix, France), ICAM-1/CD54 (BD Biosciences, Le Pont de Claix, France), and Platelet endothelial cell adhesion molecule PECAM1/CD31 (BD Biosciences, Le Pont de Claix, France). The cells were incubated for 30 minutes with antibodies. In all experiments, background labelling was assessed using the relevant fluorochrome-conjugated mouse IgG isotype control (BD Biosciences, Le Pont de Claix, France). Cells were centrifuged at 300 g for 5 minutes at 22°C then washed in PBS. Data acquisition was performed using a Guava easyCyte HT Flow Cytometer (Merck Millipore, Molsheim, France) and analysis carried out using the Incyte program. At least 10,000 events were collected for each sample.
3
Endothelial Cell Surface Marker Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The marker surface expression of EA.hy926 endothelial cells was determined by flow cytometry. Cells were fixed beforehand as described above. Direct labelling was performed for the following markers: endoglin-CD105 (BD Biosciences Cat# 563264, RRID:AB_2738104 ), ICAM-1-CD54 (BD Biosciences Cat# 740978, RRID:AB_2740602 ) and Platelet endothelial cell adhesion molecule PECAM1-CD31 (BD Biosciences Cat# 744077, RRID:AB_2741979 ). CD54 and CD31 were two principal adhesion molecules, which determine changes in endothelial permeability, activation and transendothelial leukocyte migration. The cells were incubated for 30 min with antibodies. In all of the experiments, background labelling was assessed using the relevant fluorochrome-conjugated mouse IgG isotype control (BD Biosciences, Le Pont de Claix, France). Cells were centrifuged at 300 g for 5 min at 22 °C and then washed in PBS. Data acquisition was performed using a Guava easyCyte HT Flow Cytometer (Merck Millipore, Molsheim, France) and the data were analysed using the Incyte programme. At least 10,000 events were collected for each sample.
Mycoplasma testing was performed to confirm the absence of contamination according to the recommended protocol (MycoStrip™–Mycoplasma Detection Kit # rep-mys-50 InvivoGen).
Mycoplasma testing was performed to confirm the absence of contamination according to the recommended protocol (MycoStrip™–Mycoplasma Detection Kit # rep-mys-50 InvivoGen).
4
Nanofibrillar Films and Atherosclerosis Resistance
5
Neutrophil Subset Characterization by Flow Cytometry
6
Characterization and Differentiation of MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunophenotypic characterization and differentiation capacities of MSCs was performed according to the methodology described by Montesinos et al. [35 (link)]. Monoclonal antibodies conjugated to FITC, PE, or APC against surface markers characteristic of MSCs (CD90, CD105, CD73, CD13, HLA-I, HLA-II, CD45, CD31, CD34, CD14, and CD54/ICAM-1 (BD Biosciences, San Diego, CA)) were used for immunophenotypic characterizations, as described in Flow Cytometry Analysis of Cells.
Adipogenic and osteogenic differentiation was induced with Stem Cell Kits (Stemcell Technologies, Inc., Vancouver, BC, Canada). Adipogenic differentiation was determined by visualizing the presence of Oil Red O-stained lipid vacuoles (Sigma-Aldrich, St. Louis, MO). Osteogenic differentiation was determined by alkaline phosphatase activity which was detected using SIGMAFAST™ BCIP/NBT (Sigma-Aldrich). Chondrogenic differentiation was induced with a commercial induction medium supplemented with 10 ng/mL of TGF-β (both reagents of Cambrex Bio Science). The resulting micromasses were fixed, dehydrated, embedded in paraffin, and sliced. Cross sections of 5 μm were stained with Alcian blue dye (Sigma-Aldrich).
Adipogenic and osteogenic differentiation was induced with Stem Cell Kits (Stemcell Technologies, Inc., Vancouver, BC, Canada). Adipogenic differentiation was determined by visualizing the presence of Oil Red O-stained lipid vacuoles (Sigma-Aldrich, St. Louis, MO). Osteogenic differentiation was determined by alkaline phosphatase activity which was detected using SIGMAFAST™ BCIP/NBT (Sigma-Aldrich). Chondrogenic differentiation was induced with a commercial induction medium supplemented with 10 ng/mL of TGF-β (both reagents of Cambrex Bio Science). The resulting micromasses were fixed, dehydrated, embedded in paraffin, and sliced. Cross sections of 5 μm were stained with Alcian blue dye (Sigma-Aldrich).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!