APC anti-mouse CD8, APC anti-mouse CD4, APC anti-mouse CD3, PE anti-mouse IFN-γ, and PE anti-mouse PD-1 antibodies (Abs) were obtained from BD Biosciences (San Jose, CA). Intracellular cytokine staining (ICS) of IFN-γ was performed as previously described23 (link) using a Cytofix/Cytoperm Plus (with GolgiStop) kit from BD Biosciences. Foxp3 staining was performed using the Foxp3 staining buffer set from eBioscience (San Diego, CA). Appropriate isotype controls were used for the flow cytometric analysis. Flow cytometric data were collected using the BD FACSCalibur flow cytometer (BD Biosciences). Data were analyzed using the Cell Quest (BD Biosciences).
Protocol detail
Multiparametric Flow Cytometry Analysis
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
APC anti-mouse CD4
APC anti-mouse CD3
PE anti-mouse IFN-γ
Foxp3 staining buffer set
BD FACSCalibur
Cell Quest
Anti antibodies
Anti cd3
Buffer
Cell
Cytokine
Flow cytometric
Ifn γ
Intracellular
Isotype
Mouse
Mouse ifn γ
Corresponding Organization : Shiga University of Medical Science
Other organizations : Sidney Kimmel Comprehensive Cancer Center
Top 5 similar protocols
Variable analysis
independent variables
- APC anti-mouse CD8 antibody
- APC anti-mouse CD4 antibody
- APC anti-mouse CD3 antibody
- PE anti-mouse IFN-γ antibody
- PE anti-mouse PD-1 antibody
- IFN-γ expression (intracellular cytokine staining)
- PD-1 expression
- Appropriate isotype controls for flow cytometric analysis
- Positive control: Not explicitly mentioned
- Negative control: Appropriate isotype controls for flow cytometric analysis
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!