Immunophenotyping was performed to determine cell surface marker profiles of hMSCs before and after gene targeting according to ISCT consensus28 (link). Cells were trypsinized and stained to analyze with flow cytometry, with the following monoclonal antibodies (and clone IDs) from BioLegend Inc. (San Diego, CA): CD105-PE/Cy7 (43A3), CD73-APC (AD2), CD90-PerCP/Cy5.5 (5E10), CD14-Pacific Blue (M5E2), CD19-Pacific Blue (SJ25C1), CD34-Pacific Blue (581), CD45-Pacific Blue (2D1), and HLA-DR-Pacific Blue (L243). All antibodies were used at 1:100 dilutions for staining.
Cd34 pacific blue
Manufactured by BioLegend
Sourced in France
CD34-Pacific Blue is a fluorochrome-conjugated antibody that binds to the CD34 protein, which is expressed on hematopoietic stem and progenitor cells. This product is designed for use in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Cd19 pacific blue, by BioLegend (1 mentions)
Cd14 pacific blue, by BioLegend (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Cd14 pe cy7, by BD (1 mentions)
Cd45 fitc, by BD (1 mentions)
Cd34 apc, by BD (1 mentions)
Cd33 pe, by BD (1 mentions)
Cd10 pe cy7, by BioLegend (1 mentions)
Cd19 pe cy7, by BioLegend (1 mentions)
Cd90 apc, by BioLegend (1 mentions)
Diva software, by BD (1 mentions)
Cd41 apc, by BioLegend (1 mentions)
Cd42 pe, by BD (1 mentions)
Cd45 alexa fluor 700, by BioLegend (1 mentions)
Facsaria 2 cell sorter, by BD (1 mentions)
Human progenitor cell enrichment kit, by STEMCELL (1 mentions)
Cd45 pe cy7, by BioLegend (1 mentions)
Cd133 pe, by BioLegend (1 mentions)
Fcs express, by De Novo Software (1 mentions)
Stemspan media, by STEMCELL (1 mentions)
4 protocols using cd34 pacific blue
1
Immunophenotyping of Gene-Targeted hMSCs
2
Multiparametric Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed in phosphate-buffered saline and incubated for 30 minutes at 4 °C with a combination of monoclonal antibodies against human cell surface antigens as presented in Table 1 . Antibodies were purchased as described: CD36 APC, CD14 PE-Cy7, CD42 PE, CD33 PE, CD34 APC, CD45 FITC (all from Becton Dickinson), CD34 Pacific Blue, CD45 Alexa Fluor 700, CD41 APC, CD90 APC, CD10 PE-Cy7, CD19 PE-Cy7 (all from Biolegend, Saint Quentin en Yvelines, France), and CD235a PE, 7-AAD (both from Beckmann Coulter, Villepinte, France). Cells were analyzed on a FACSCanto II flow cytometer (Becton Dickinson) using Diva software (version 6.1.3; Becton Dickinson).
3
Isolation of Progenitor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mononuclear cell fractions from healthy individuals or patients with diabetes were negatively selected for lineage-negative (Lin−) population using a human progenitor cell enrichment kit (Stem Cell Technologies, Vancouver, BC, Canada). Lin− cells were stained with a Pacific Blue CD34 and PE/Cy7 CD45 antibodies (BioLegend, San Diego, CA) and sorted for Lin−CD34highCD45dim population using a FACSAria II cell sorter (BD Biosciences, San Jose, CA).
4
Characterization of Hematopoietic Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isolated CD34+ cells were propagated in round-bottom 96-well culture plates with no more than 5,000 cells per well under two different culturing conditions for 4 days: 1) undifferentiated, Stem Span Media supplemented with cytokine cocktail containing IL-3, IL-6, Flt, and SCF (Stem Cell Technologies) and 2) differentiated, EBM-2 media supplemented with EGM-2MV growth supplements (Lonza, Allendale, NJ) containing human epidermal growth factor, hydrocortisone, human fibroblast growth factor beta, VEGF, recombinant analog of insulin-like growth factor, ascorbic acid, heparin, and FBS.
At 4 days, the cells were stained with a cocktail of antibodies containing Pacific Blue CD34, PE/Cy7 CD45, FITC CD144, and PE CD133 (BioLegend). Cell-surface expression was analyzed using a BD LSR II Flow Cytometer, and the analysis was performed using FCS express (De Novo Software, Los Angeles, CA).
At 4 days, the cells were stained with a cocktail of antibodies containing Pacific Blue CD34, PE/Cy7 CD45, FITC CD144, and PE CD133 (BioLegend). Cell-surface expression was analyzed using a BD LSR II Flow Cytometer, and the analysis was performed using FCS express (De Novo Software, Los Angeles, CA).
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!