Evaluation of AnxA2 and Its Phosphorylation

Elution of AnxA2 (26 (link))and Western blot with rabbit anti-AnxA2, mouse anti-P-Y23-AnxA2, mouse anti-beta-actin, rabbit anti-HGF (all from Santa Cruz Biotechnology), rabbit anti-IGF-1(Abcam) or goat anti-Shh (R&D Systems) antibodies were described previously (12 (link)) Immunohistochemistry (IHC) staining for E-Cadherin and HGF was performed with rabbit anti-E-Cadherin (Abcam) and rabbit anti-HGF (Santa Cruz Biotechnology) antibodies using a standard protocol on an automated stainer from Leica Microsystems. IHC for SMA was performed as previously described (27 (link)). IHC for AnxA2 was conducted with mouse anti-AnxA2(Invitrogen), or mouse anti-P-Y23-AnxA2 antibodies, IHC for Shh with goat-anti-Shh (R&D), and IHC for IGF-1 with rabbit-anti-IGF-1(Abcam) antibodies as described previously (13 (link)). All IHC slides were analyzed and scored by a pathologist (A.L). The levels of secreted IGF-1 were measured by ELISA (R&D) following the manufacturer’s instructions.

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.

Rucki A.A., Foley K., Zhang P., Xiao Q., Kleponis J., Wu A.A., Sharma R., Mo G., Liu A., Van Eyk J., Jaffee E.M, & Zheng L. (2016). Heterogeneous stromal signaling within the tumor microenvironment controls the metastasis of pancreatic cancer. Cancer research, 77(1), 41-52.

Publication 2016

rabbit anti-AnxA2 mouse anti-beta-actin rabbit anti-HGF rabbit anti-IGF-1 goat anti-Shh rabbit anti-E-Cadherin automated stainer

Anti p antibodies Antibodies Anxa2 Beta actin Cadherin Elisa Goat Igf 1 Immunohistochemistry Mouse Pathologist Rabbit Western blot

Corresponding Organization : Sidney Kimmel Comprehensive Cancer Center

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Elution of AnxA2
Western blot with rabbit anti-AnxA2, mouse anti-P-Y23-AnxA2, mouse anti-beta-actin, rabbit anti-HGF, rabbit anti-IGF-1, or goat anti-Shh antibodies
IHC staining for E-Cadherin and HGF with rabbit anti-E-Cadherin and rabbit anti-HGF antibodies
IHC for SMA as previously described
IHC for AnxA2 with mouse anti-AnxA2 or mouse anti-P-Y23-AnxA2 antibodies
IHC for Shh with goat-anti-Shh antibody
IHC for IGF-1 with rabbit-anti-IGF-1 antibody

dependent variables

Levels of secreted IGF-1 measured by ELISA

control variables

Staining protocol on an automated stainer from Leica Microsystems for IHC
Previously described protocol for IHC for SMA

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!