Embryonic and Postnatal Tissue Immunohistochemistry

Embryonic brains, skin, heart or lungs were dissected from pregnant females at E15.5 and E17.5 and postnatal brains from P0 and P7 pups. P22 mice were euthanized with isoflurane overdose and the brains were perfused with 4% paraformaldehyde (PFA). All other brains and organs were drop fixed in 4% PFA overnight. Immunohistochemistry was carried out essentially as described previously on 50 μm-thick Vibratome sections (Ma et al. 2012 (link), 2013 (link)). Primary antibodies or reagents used include biotinylated isolectin B4 (Vector Lab), rat anti-mouse CD31 (BD Pharmingen), rabbit anti-GFP (Life Technologies), rabbit anti-Brain lipid binding protein (Millipore), rabbit anti-Desmin (Millipore), mouse anti-NeuN (Millipore) and rat anti-mouse endomucin (Affymetrix eBioscience). FITC or Cy3 conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA). Images were taken under a Nikon eclipse Ti microscope and processed using Adobe Photoshop CS6.

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.

Santhosh D, & Huang Z. (2016). A Tie2-driven BAC-TRAP transgenic line for in vivo endothelial gene profiling. Genesis (New York, N.Y. : 2000), 54(3), 136-145.

Publication 2016

biotinylated isolectin B4 rat anti-mouse CD31 rabbit anti-GFP mouse anti-NeuN FITC or Cy3 conjugated secondary antibodies

Antibodies Brain lipid binding protein Brains Desmin Embryonic Endomucin Fitc Heart Immunohistochemistry Isoflurane Isolectin Lungs Microscope Mouse Overdose Paraformaldehyde Pregnant females Rabbit Skin Vector

Corresponding Organization : University of Wisconsin–Madison

Top 5 similar protocols

Variable analysis

independent variables

Developmental stage (E15.5, E17.5, P0, P7, P22)

dependent variables

Expression of biotinylated isolectin B4, CD31, GFP, Brain lipid binding protein, Desmin, NeuN, and endomucin

control variables

Tissue preparation (dissection, fixation, and sectioning)
Immunohistochemistry protocol

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly 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!