Dual Labeling of CRFR1-GFP Neurons

Complete series through the brains of 4 CRFR1-GFP mice were prepared for concurrent localization of GFP and CRFR1 mRNA. Combining immunoperoxidase labeling with isotopic in situ hybridization required minor modifications of the constituent methods (Chan et al., 1993 (link)). Immunostaining was carried out first, and the protocols modified as follows: (1) normal tissue pretreatments in hydrogen peroxide and sodium borohydride were omitted, (2) blocking sera were replaced in the immunostaining procedure with 2% bovine serum albumin and 2% heparin sulfate, (3) nickel enhancement steps were eliminated, and (4) Nissl counterstaining was omitted.

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.

Justice N.J., Yuan Z.F., Sawchenko P.E, & Vale W. (2008). Type 1 Corticotropin-Releasing Factor Receptor Expression Reported in BAC Transgenic Mice: Implications for Reconciling Ligand-Receptor Mismatch in the Central CRF System. The Journal of comparative neurology, 511(4), 479-496.

Publication 2008

Bovine serum albumin Brains Heparin Hydrogen peroxide Immunoperoxidase In situ hybridization Mice Mrna Nickel Normal tissue Sera Sodium borohydride Sulfate

Corresponding Organization :

Other organizations : Salk Institute for Biological Studies, Tzu Chi University

Top 5 similar protocols

Protocol cited in 15 other protocols

Variable analysis

independent variables

Modifications to the immunostaining protocol:

dependent variables

Concurrent localization of GFP and CRFR1 mRNA

control variables

Immunostaining protocol from Chan et al. (1993) as the control method

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!