After viability determination of harvested lymphocytes from each group using 0.04% trypan blue (viability > 90%) and adjusted cell concentration to 1 × 106 cells/ml in PBS containing 2% FBS, the cells were incubated with surface markers including phycoerythrin (PE)-labeled anti-mouse CD3, Allophycocyanin (APC)-labeled anti-mouse CD4 and fluorescein isothiocyanate (FITC)-labeled anti-mouse CD8 (eBioscience) at 4°C for 30 min in the dark. Followed by washed by 2 ml PBS and then fixed with FACScan buffer (PBS containing 1% FCS and 0.1% Sodium azide) and 2% paraformaldehyde, the cultures were analyzed of fluorescence profiles on a FACScan flow cytometer (BD Bio-sciences) by SYSTEM II software (Coulter).
Pe labeled anti mouse cd3
Manufactured by Thermo Fisher Scientific
Sourced in United States
PE)-labeled anti-mouse CD3 is a laboratory reagent used for the identification and analysis of mouse CD3+ T cells. It is a monoclonal antibody conjugated with the fluorescent dye R-Phycoerythrin (PE), which binds specifically to the CD3 surface antigen expressed on mouse T cells. This product can be used in flow cytometry and other immunoassays to detect and quantify mouse T cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Allophycocyanin apc labeled anti mouse cd4, by Thermo Fisher Scientific (5 mentions)
Facscan flow cytometer, by BD (5 mentions)
Fluorescein isothiocyanate fitc labeled anti mouse cd8, by Thermo Fisher Scientific (4 mentions)
System 2 software, by Beckman Coulter (3 mentions)
Fitc labeled anti mouse cd8, by Thermo Fisher Scientific (3 mentions)
Apc labeled anti mouse cd4, by Thermo Fisher Scientific (2 mentions)
Facscan, by BD (1 mentions)
7 protocols using pe labeled anti mouse cd3
1
Flow Cytometry Analysis of Immune Cells
2
Quantifying T cell subsets by flow cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentration of the purified splenocytes were adjusted into 1× 105 cells/mL. Then the phycoerythrin (PE)-labeled anti-mouse CD3 (eBioscience) (5 μg/mL), Allophycocyanin (APC)-labeled anti-mouse CD4 (eBioscience) (5 μg/mL) and fluorescein isothiocyanate (FITC)-labeled anti-mouse CD8 (eBioscience) (5 μg/mL) antibodies were used to stain the T cell subclasses (CD4+ and CD8+) for 30 min at 4 °C. After washing by PBS, the cultures were fixed with FACScan buffer (1% FCS and 0.1% Sodium azide in PBS) and 2% paraformaldehyde. The T cell subclasses were measured for fluorescence profiles on a FACScan flow cytometer (BD Bio-sciences) and analyzed by SYSTEM II software (Coulter).
3
Flow Cytometric Analysis of T Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Splenic lymphocytes were harvested as described above, viability was determined using 0.04% trypan blue (viability > 90%) and cell concentration was adjusted to 1 × 106 cells/mL in PBS containing 2% FBS. As in a previous study [37 (link)], the lymphocytes were incubated with surface markers including phycoerythrin (PE)-labeled anti-mouse CD3, allophycocyanin (APC)-labeled anti-mouse CD4 and fluorescein isothiocyanate (FITC)-labeled anti-mouse CD8 (eBioscience) at 4°C for 30 min in the dark. Then, the cells were fixed with FACScan buffer (PBS containing 1% FCS, 0.1% sodium azide and 2% paraformaldehyde). The analysis of surface markers (CD3, CD4 and CD8) on the cells was performed with fluorescence profiles through a FACScan flow cytometer (BD Biosciences, USA).
4
Quantifying T Cell Subsets in Spleen
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The percentage of T cell subclasses including CD4+ and CD8+ in spleen were determined by flow cytomety analysis. With staining by phycoerythrin (PE)-labeled anti-mouse CD3 (eBioscience, San Diego, California, USA), allophycocyanin (APC)-labeled anti-mouse CD4 (eBioscience) and fluorescein isothiocyanate (FITC)-labeled anti-mouse CD8 (eBioscience) antibodies, the single-cell suspension splenocytes were washed by PBS, the cells were then fixed with FACScan buffer (PBS containing 1% FCS and 0.1% Sodium azide) and 2% paraformaldehyde. All samples were analyzed using SYSTEM II software through FACScan flow cytometer (BD Biosciences, San Jose, California, USA).
5
Lymphocyte Surface Marker Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lymphocytes were isolated from spleen, as described above. According to the method described in our previous studies [3 (link), 40 (link)], surface staining was performed by incubation with surface markers, including phycoerythrin-(PE)-labeled anti-mouse CD3, allophycocyanin (APC)-labeled anti-mouse CD4, and fluorescein isothiocyanate (FITC)-labeled anti-mouse CD8 (eBioscience). The cell suspension was then fixed with FACScan buffer (PBS containing 1% BSA and 0.1% sodium azide) and 2% paraformaldehyde. All data were acquired through a FACScan flow cytometer (BD Biosciences, USA). The analysis was performed with the data from three independent experiments.
6
Flow Cytometric Analysis of T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Analyses of CD4+ and CD8+ T lymphocytes were performed according to our previous study [25 (link)]. The percentages of CD4+ and CD8+ T lymphocytes were determined using flow cytometry with staining by the surface markers including phycoerythrin- (PE-) labeled anti-mouse CD3 (eBioscience), allophycocyanin- (APC-) labeled anti-mouse CD4 (eBioscience), and fluorescein isothiocyanate- (FITC-) labeled anti-mouse CD8 (eBioscience) antibodies. All the samples were analyzed regarding fluorescence profiles on a FACScan flow cytometer (BD Biosciences) by SYSTEM II software (Coulter).
7
Multicolor Flow Cytometry Immunophenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The splenic lymphocytes were collected as described above [18] ,
The viability was determined with 0.04% trypan blue (viability > 90%) and alive cell concentration was adjusted to 1 × 10 6 cells/ml in PBS containing 2% FBS, and the lymphocytes were incubated with 3 different surface markers at 4 °C for 30 min in the dark [19, 20] , including phycoerythrin (PE)-labeled anti-mouse CD3, allophycocyanin (APC)-labeled antimouse CD4 and fluorescein isothiocyanate (FITC)labeled anti-mouse CD8 (eBioscience). Then the cells were fixed with FACScan buffer (PBS containing 1% FCS and 0.1% Sodium azide) and 2% paraformaldehyde. The specific cells were analyzed according to the surface markers (CD3, CD4 and CD8) through a FACScan flow cytometer (BD Biosciences, USA).
The viability was determined with 0.04% trypan blue (viability > 90%) and alive cell concentration was adjusted to 1 × 10 6 cells/ml in PBS containing 2% FBS, and the lymphocytes were incubated with 3 different surface markers at 4 °C for 30 min in the dark [19, 20] , including phycoerythrin (PE)-labeled anti-mouse CD3, allophycocyanin (APC)-labeled antimouse CD4 and fluorescein isothiocyanate (FITC)labeled anti-mouse CD8 (eBioscience). Then the cells were fixed with FACScan buffer (PBS containing 1% FCS and 0.1% Sodium azide) and 2% paraformaldehyde. The specific cells were analyzed according to the surface markers (CD3, CD4 and CD8) through a FACScan flow cytometer (BD Biosciences, USA).
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!