Comprehensive Tissue Analysis Protocol

Histological and immunohistochemical analyses were performed as previously described^{2 (link)}. The following antibodies were used: Cdh1 (rat, Novex), Cortactin (rabbit, Abcam), GFP (goat, Abcam), GFP (mouse, Roche), ItgA2 (rabbit, Abcam), ItgA6 (rabbit, Abcam), ItgB1 (rabbit, Abcam), Ki67 (rat, Biolegend), Krt19 TROMA-III (rat, DSHB), MLC2 pSer19 (rabbit, NEB), Myosin (rabbit, Abcam), Nestin (mouse, BD transduction), Pdgfrβ (rabbit, NEB), PTK2 pTyr397 (rabbit, ThermoFisher), SMA (mouse, Agilent), SMA (mouse, Sigma), Tomato (rabbit, Rockland), Tomato (goat, Biorbyt), Vimentin (rabbit, NEB), Vinculin (mouse, Sigma-Aldrich). DBA-rhodamine and DBA-FITC were from Vectorlabs. F-actin was stained with Phalloidin-TRITC (Sigma-Aldrich) and nuclei with DAPI (Sigma-Aldrich). F-actin staining of LSL-KrasG12D; p53 F/F; Pdx1-Cre pancreata and wildtype littermate control pancreata was performed on cryosections. Samples were embedded fresh in OCT medium and after sectioning fixed in 5% NBF for 10 min. Slides were washed in 0.2% Triton X-100 in PBS for 10 min and incubated in FLASH blocking buffer for 30 min. Staining reagent incubations were performed as above. Fluorescent stainings were imaged on a Zeiss LSM 780 confocal microscope. Chromogenic DAB stainings were imaged on a Zeiss Axio Scan Z1 Slide Scanner.

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.

Messal H.A., Alt S., Ferreira R.M., Gribben C., Wang V.M., Cotoi C., Salbreux G, & Behrens A. (2019). Tissue curvature and apicobasal mechanical tension imbalance instruct cancer morphogenesis. Nature, 566(7742), 126-130.

Publication 2019

Cortactin ItgA2 ItgA6 ItgB1 Myosin Nestin Vinculin DBA-rhodamine DBA-FITC Phalloidin-TRITC DAPI LSM 780 confocal microscope Axio Scan Z1 Slide Scanner

Antibodies Buffer Cdh1 Chromogenic Confocal microscope Cortactin Cryosections Dapi F actin Fitc Goat Krt19 Mouse Myosin Nestin Nuclei Pancreata Pdgfrβ Pdx1 Ptk2 Rabbit Rhodamine Scan Stainings Tomato Tritc phalloidin Triton x 100 Vimentin Vinculin

Corresponding Organization : King's College London

Other organizations : The Francis Crick Institute, King's College Hospital

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Antibodies used for immunohistochemical analyses: Cdh1, Cortactin, GFP, ItgA2, ItgA6, ItgB1, Ki67, Krt19 TROMA-III, MLC2 pSer19, Myosin, Nestin, Pdgfrβ, PTK2 pTyr397, SMA, Tomato, Vimentin, Vinculin

dependent variables

Histological and immunohistochemical analyses

control variables

Samples were embedded fresh in OCT medium and after sectioning fixed in 5% NBF for 10 min.
Slides were washed in 0.2% Triton X-100 in PBS for 10 min and incubated in FLASH blocking buffer for 30 min.
Staining reagent incubations were performed as previously described.
Fluorescent stainings were imaged on a Zeiss LSM 780 confocal microscope.
Chromogenic DAB stainings were imaged on a Zeiss Axio Scan Z1 Slide Scanner.

controls

DBA-rhodamine and DBA-FITC were used as staining reagents.
F-actin was stained with Phalloidin-TRITC and nuclei with DAPI.

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!