Comprehensive Murine Tissue Analysis Panel

For flow cytometry and imaging studies with murine tissues, the following antibodies were purchased from Biolegend: anti-CD45 (30-F11), anti-CD31 (390), anti-PDPN (8.1.1), anti-α-smooth muscle actin (1A4), anti-Thy1 (53–2.1), anti-CD140α (APA5), anti-CD140b (APB5), anti-CD106 (429), anti-CD4 (RM4.5), and anti-CD8 (RM2206). The anti-FAP used for flow cytometry analysis was provided by Ellen Pure and has been validated (anti-mouse FAP 73.3)(8 (link)). For immunofluorescent analysis, the Roche anti-FAP 28H1 clone was used (26 (link)). Functional-grade purified anti-CD3ε (145–2C11) was from BD Biosciences, and anti-CD28 (37.51) was from Biolegend. Antibodies used to stain human samples include viability dye (L10119; Life Technologies), anti-PDPN (clone NZ1.3; eBioscience), anti-Thy1 (clone 5E10; BD Biosciences), anti-CD4 (clone A161A1; Biolegend) and anti-CD8 (clone HIT8α; Biolegend). For PBMC stimulation, anti-human CD3 (clone OKT3; eBioscience) and anti-human CD28 (clone CD28.2; eBioscience) were used. L-NMMA (NG-monomethyl-L-arginine) was purchased from Calbiochem and CFSE (carboxyfluorescein diacetate succinimidyl ester) was from Invitrogen.

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.

Cremasco V., Astarita J., Grauel A.L., Keerthivasan S., MacIsaac K., Woodruff M.C., Wu M., Spel L., Santoro S., Amoozgar Z., Laszewski T., Migoni S.C., Knoblich K., Fletcher A.L., LaFleur M., Wucherpfennig K.W., Pure E., Dranoff G., Carroll M.C, & Turley S.J. (2018). FAP delineates heterogeneous and functionally divergent stromal cells in immune-excluded breast tumors. Cancer immunology research, 6(12), 1472-1485.

Publication 2018

anti-CD8 (RM2206). anti-CD3ε (145–2C11) anti-CD28 (37.51) anti-PDPN (clone NZ1.3; anti-Thy1 (clone 5E10; anti-CD4 (clone A161A1; anti-human CD3 (clone OKT3; anti-human CD28 (clone CD28.2; CFSE

Actin Antibodies Carboxyfluorescein diacetate succinimidyl ester Cd3ε Clone Flow cytometry Human Immunofluorescent Mouse Ng monomethyl l arginine Okt3 Smooth muscle Stain Tissues

Corresponding Organization : Harvard University

Other organizations : Boston Children's Hospital, University of Birmingham, Australian Regenerative Medicine Institute, Monash University, University of Pennsylvania

Top 5 similar protocols

Variable analysis

independent variables

Anti-CD3ε (145–2C11)
Anti-CD28 (37.51)
Anti-human CD3 (clone OKT3)
Anti-human CD28 (clone CD28.2)

dependent variables

Flow cytometry and imaging studies with murine tissues
Immunofluorescent analysis
PBMC stimulation

control variables

Murine tissues
Human samples
PBMCs
Viability dye (L10119; Life Technologies)
L-NMMA (NG-monomethyl-L-arginine)
CFSE (carboxyfluorescein diacetate succinimidyl ester)

positive controls

Anti-mouse FAP 73.3 (validated)
Roche anti-FAP 28H1 clone

negative controls

No negative controls 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!