Immunoblotting and Immunofluorescence Analysis

Immunoblotting was performed as described previously [44 (link)]. Equal amounts of tissue lysates were used for immunoblotting. Blots were incubated with specific antibodies to BMP-1 and latent TGFβ1-binding protein-1 (LTBP-1) (ab205394 and ab78294; Abcam), Flag (F3165; Sigma-Aldrich), ALK5 (MAB5871; R&D Systems), Col3a1 and Fn1 (NB600 and NBP1-91258; Novus Biologicals). β-Actin (A2228; Sigma-Aldrich) was used as a loading control. Immunofluorescence was performed as described in detail previously [44 (link)]. We used specific antibodies to CD34 (553731; BD Bioscience or ab54208; Abcam), Nkx2.1 (ab76013; Abcam) and VE-cadherin (562243; BD Biosciences). The nuclei were stained with 4′,6-diamidino-2-phenylindole (D9564; Sigma-Aldrich).

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.

Wu X., Zhang D., Qiao X., Zhang L., Cai X., Ji J., Ma J.A., Zhao Y., Belperio J.A., Boström K.I, & Yao Y. (2023). Regulating the cell shift of endothelial cell-like myofibroblasts in pulmonary fibrosis. The European Respiratory Journal, 61(6), 2201799.

Publication 2023

ab205394 ab78294 F3165 MAB5871 NB600 NBP1-91258 β-Actin A2228 ab54208 ab76013 4′,6-diamidino-2-phenylindole D9564

Actin Alk5 Antibodies Biologicals Bmp 1 Immunofluorescence Nkx2 1 Novus Nuclei Protein 1 Tgfβ1 Tissue Ve cadherin

Corresponding Organization :

Other organizations : University of California, Los Angeles, Stanford University

Top 5 similar protocols

Variable analysis

independent variables

Immunoblotting was performed with specific antibodies to BMP-1 and latent TGFβ1-binding protein-1 (LTBP-1), Flag, ALK5, Col3a1, and Fn1
Immunofluorescence was performed with specific antibodies to CD34, Nkx2.1, and VE-cadherin

dependent variables

Tissue lysate expression levels of BMP-1, latent TGFβ1-binding protein-1 (LTBP-1), Flag, ALK5, Col3a1, and Fn1
Cellular localization and expression of CD34, Nkx2.1, and VE-cadherin

control variables

β-Actin was used as a loading control for immunoblotting
The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) for immunofluorescence

controls

Positive control: Not specified
Negative control: Not specified

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!