PBMCs were isolated according to methods described elsewhere (16 (link)). PBMCs were blocked using a rat anti-mouse CD16/32 antibody (BD Pharmingen) and stained with FITC conjugated anti-mouse CD45.2 (clone: 104), APC-eFluor780 conjugated anti-mouse CD45.1 (clone: A20), PE conjugated anti-mouse B220 (clone: RA3–6B2), PE-Cy5 conjugated anti-mouse CD11b (clone: M1/70) (all acquired from eBioscience) and Brilliant Violet 421 conjugated anti-mouse CD3e antibodies (clone: 145–2C11; BioLegend). All antibodies were diluted 1:400. Data were collected from at least 20,000 single cells by FACSCanto (BD Pharmingen) and analyzed using FlowJo (Tree Star, Inc) without knowledge of the genotype or treatment by a single observer (C-LL).
Protocol detail
Immunophenotyping of Mouse PBMCs
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.
Corresponding Organization :
Other organizations : Duke University Hospital, Duke Medical Center
Top 5 similar protocols
Variable analysis
independent variables
- Genotype
- Treatment
- Percentage of CD45.2+ cells
- Percentage of CD45.1+ cells
- Percentage of B220+ cells
- Percentage of CD11b+ cells
- Percentage of CD3e+ cells
- Cell isolation method (described elsewhere in reference 16)
- Blocking with rat anti-mouse CD16/32 antibody
- Antibody concentrations (all diluted 1:400)
- Data collection (at least 20,000 single cells)
- Data analysis (performed by a single observer without knowledge of genotype or treatment)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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!