To characterise the MSCs, they were washed twice in phosphate-buffered saline (PBS; Sigma-Aldrich, St. Louis, MO, USA) then pre-blocked with 2% rat serum in PBS. The following direct antibodies were used: phycoerythrin (PE)-conjugated mouse anti-human CD34 (1:20; DakoCytomation, Barcelona, Spain), fluorescein isothiocyanate (FITC)-conjugated mouse anti-rat CD45 (1:20; BD Pharmingen, Franklin Lakes, NJ, USA), PE-cyanine (Cy)5.5-conjugated mouse anti-rat CD90 (1:20; Immunostep, Salamanca, Spain) and allophycocyanin (APC)-conjugated mouse anti-rat CD29 (1:20; Immunostep, Salamanca, Spain). The cells were washed with PBS after 1 h of incubation with the corresponding antibody at room temperature. Fluorescence-activated cell sorting (FACS) data was generated by BD FACSDiva software (BD Science, San Jose, CA, USA). Negative control staining was performed using FITC-conjugated mouse IgG1K isotype, PE-conjugated mouse IgG1K isotype, PE-Cy5.5-conjugated mouse IgG1K isotype and APC-conjugated mouse IgG1K isotype (BD Pharmingen, Franklin Lakes, NJ, USA).
Fitc conjugated mouse igg1k isotype
Manufactured by BD
Sourced in United States, Spain
FITC-conjugated mouse IgG1K isotype is a laboratory reagent used for various immunological assays. It consists of a fluorescein isothiocyanate (FITC) molecule conjugated to a mouse immunoglobulin G1 (IgG1) antibody of the kappa (K) light chain isotype. This reagent can be used to detect and quantify target antigens in samples such as cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Pe conjugated mouse igg1k isotype, by BD (5 mentions)
Pe cy5.5 conjugated mouse igg1k isotype, by BD (5 mentions)
Apc conjugated mouse igg1k isotype, by BD (3 mentions)
Diva software, by BD (3 mentions)
Facsaria flow cytometer, by BD (2 mentions)
Fitc mouse anti human cd105, by Bio-Rad (2 mentions)
Fitc mouse anti human cd45, by BD (2 mentions)
Pe cy5.5 conjugated mouse anti human cd90, by BD (2 mentions)
Pe conjugated anti human cd73, by BD (2 mentions)
Facsdiva software, by BD (1 mentions)
Facsaria, by BD (1 mentions)
Attune nxt, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Novoexpress software, by Agilent Technologies (1 mentions)
5 protocols using fitc conjugated mouse igg1k isotype
1
Characterization of Mesenchymal Stem Cells
2
Characterization of MSCs Across Ages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To characterize the different populations of MSCs from chronologically different animals, their MSCs were washed twice in PBS, then pre-blocked with 2% rat serum in PBS. The following direct antibodies were used: PE-conjugated mouse anti-rat CD34 (1:20 from DakoCytomation, Barcelona, SP); FITC-conjugated mouse anti-rat CD45 (1:20 BD Pharmingen, New Jersey, USA); PE-Cy5.5-conjugated mouse anti-rat CD90 (1:20 Immunostep, Salamanca, SP) and APC-conjugated mouse anti-rat CD29 (1:20 Immunostep, Salamanca, SP). The cells were washed with PBS after one hour of incubation with the corresponding antibody at room temperature. The stained cells were then washed twice with PBS, and 2 × 105 cells were analysed with a FACSAria flow cytometer (BD Science, Madrid, SP). FACS data were generated by DIVA software (BD Science). Negative control staining was performed using FITC-conjugated mouse IgG1K isotype, PE-conjugated mouse IgG1K isotype, PE-Cy5.5-conjugated mouse IgG1K isotype and APC-conjugated mouse IgG1K isotype (all from BD Pharmingen).
3
Multicolor Flow Cytometry of Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were washed twice with PBS and incubated for 1 h at RT with the following direct antibodies: phycoerythrin (PE) mouse anti-human CD34 (1:20; DakoCytomation); FITC mouse anti-human CD45 (1:20; BD Pharmingen); FITC mouse anti-human CD105 (1:100; Serotec); PE-Cy5.5-conjugated mouse anti-human CD90 (1:20; BD Pharmingen); PE-conjugated anti-human CD73 (1:20; BD Pharmingen). The stained cells were then washed twice with PBS and 10 × 105 cells were analyzed with a FACSAria flow cytometer (BD Bioscience). FACS data were generated by DIVA software (BD Bioscience). Negative control staining was performed using FITC-conjugated mouse IgG1k isotype, PE-conjugated mouse IgG1k isotype, and PE-Cy5.5-conjugated mouse IgG1k isotype (all from BD Pharmingen).
4
Apoptosis Analysis of HGPS Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flow cytometry was performed using Attune NxT (Thermo Fisher, Brussels, Belgium) equipment to study the apoptosis potential of HGPS cell lines and their healthy progenitors. Cells were washed twice in PBS (Sigma-Aldrich, St. Louis, MO, USA) and incubated with 1 μM staurosporine (STS), 1 μM Tg, 200 μM H2O2, 60 μM menadione, or 1 μM DMSO for 5 h prior to the apoptosis analysis. Incubation with FITC -7-AAD and PE-Cy5.5-annexin V was performed to check the potential apoptosis involved with mitochondrial membrane potential and oxidative stress. Then, 2 × 105 cells were analyzed using FlowJo software V8.6 (Tree Star, San Carlos, CA, USA). Mitochondrial ROS was measured after the incubation of cells with 5 mM of Mitosox-A. The stained cells were then washed twice with PBS, and 2 × 105 cells were analyzed using NovoExpress Software (Acea Biosciences, San Diego, CA, USA). For negative control staining, we used FITC-conjugated mouse IgG1K isotype, PE-conjugated mouse IgG1K isotype, PE-Cy5.5-conjugated mouse IgG1K isotype, and APC-conjugated mouse IgG1K isotype (all from BD Pharmingen, Madrid, Spain).
5
Immunophenotyping of Human Mesenchymal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed with PBS twice then pre-blocked with 3% bovine serum albumin (BSA), or 2-5% serum from the same species, in PBS. The following direct antibodies were used: phycoerythrin (PE) mouse anti-human CD34 (1:20 from DakoCytomation, Barcelona, SP); FITC mouse anti-human CD45 (1:20 BD Pharmingen, Madrid, SP); FITC mouse anti-human CD105:
(1:100 from Serotec, Bavaria, Germany): PE-Cy5.5-conjugated mouse anti-human CD90 (1:20 from BD Pharmingen); PE-conjugated anti-human CD73 (1:20 from BD Pharmingen); and rabbit anti-human VEGF (1:20 from DakoCytomation). For detection of primary antibodies not directly labelled, the cells were washed with PBS, then incubated with polyclonal rabbit anti-mouse IgD/FITC Rabbit F(ab́) 2 (1:1,000 from DakoCytomation) for 30 min at room temperature. The stained cells were then washed twice with PBS and 1 × 10 5 cells were analyzed with a FACSAria flow cytometer (BD Bioscience, Madrid, SP). FACS data were generated by DIVA software (BD Bioscience). Negative control staining was performed using FITC-conjugated mouse IgG1k isotype, PE-conjugated mouse IgG1k isotype and PE-Cy5.5-conjugated mouse IgG1k isotype (all from BD Pharmingen).
(1:100 from Serotec, Bavaria, Germany): PE-Cy5.5-conjugated mouse anti-human CD90 (1:20 from BD Pharmingen); PE-conjugated anti-human CD73 (1:20 from BD Pharmingen); and rabbit anti-human VEGF (1:20 from DakoCytomation). For detection of primary antibodies not directly labelled, the cells were washed with PBS, then incubated with polyclonal rabbit anti-mouse IgD/FITC Rabbit F(ab́) 2 (1:1,000 from DakoCytomation) for 30 min at room temperature. The stained cells were then washed twice with PBS and 1 × 10 5 cells were analyzed with a FACSAria flow cytometer (BD Bioscience, Madrid, SP). FACS data were generated by DIVA software (BD Bioscience). Negative control staining was performed using FITC-conjugated mouse IgG1k isotype, PE-conjugated mouse IgG1k isotype and PE-Cy5.5-conjugated mouse IgG1k isotype (all from BD Pharmingen).
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!