Protocol detail

Comprehensive BM Cell Immunophenotyping

Find Similar Protocols

Whole BM cells were isolated from femurs and tibias according to methods described elsewhere (16 (link)). A total of 3 × 10⁶ live BM cells were blocked using a rat anti-mouse CD16/32 antibody (BD Pharmingen) and stained with PE-Cy5 conjugated anti-mouse CD3e (clone: 145–2C11), PE-Cy5 conjugated anti-mouse CD4 (clone: GK1.5), PE-Cy5 conjugated anti-mouse CD8 (clone: 53–6.7) PE-Cy5 conjugated anti-mouse B220 (clone: RA3–6B2), PE-Cy5 conjugated PE-Cy5 conjugated anti-mouse CD11b (clone: M1/70), PE-Cy5 conjugated anti-mouse Gr-1 (clone: RB6–8C5), PE-Cy5 conjugated anti-mouse Ter119 (Ly-76), PE conjugated anti-mouse Sca1 (clone: D7), APC conjugated anti-mouse cKit (clone: 2B8), FITC conjugated anti-mouse CD45.2 (clone: 104; eBioscience) and APC-eFluor780 conjugated anti-mouse CD45.1 (clone: A20; e eBioscience). All antibodies were diluted 1:400. Data were collected from at least 500,000 single cells using FACSCanto (BD Pharmingen) and analyzed using FlowJo (Tree Star Inc.) without knowledge of the genotype or treatment by a single observer (C-LL).

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.

Hasapis S., Caraballo I, & Lee C.L. (2021). Transplantation of Unirradiated Bone Marrow Cells after Total-Body Irradiation Prevents the Development of Thymic Lymphoma in Mice through Niche Competition. Radiation research, 195(3), 301-306.

Publication 2021

rat anti-mouse CD16/32 antibody PE-Cy5 conjugated anti-mouse CD3e PE-Cy5 conjugated anti-mouse CD4 PE-Cy5 conjugated anti-mouse CD8 PE-Cy5 conjugated anti-mouse B220 PE-Cy5 conjugated anti-mouse Gr-1 PE conjugated anti-mouse Sca1 APC conjugated anti-mouse cKit FITC conjugated anti-mouse CD45.2 FACSCanto

Anti antibody Antibodies Cd11b Cells Ckit Clone Femurs Fitc Genotype Mouse Sca1 Tibias Tree

Top 5 similar protocols

Variable analysis

independent variables

Genotype
Treatment

dependent variables

Immune cell populations measured by flow cytometry: CD3e+ (T cells), CD4+ (helper T cells), CD8+ (cytotoxic T cells), B220+ (B cells), CD11b+ (myeloid cells), Gr-1+ (granulocytes), Ter119+ (erythroid cells), Sca1+ (hematopoietic stem/progenitor cells), cKit+ (hematopoietic stem/progenitor cells)
Percentage of CD45.1+ and CD45.2+ cells

control variables

Bone marrow cell isolation method
Flow cytometry staining protocol
Flow cytometry data analysis by a single observer without knowledge of genotype or treatment

controls

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.

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?

Revolutionizing how scientists
search and build protocols!