Podocyte Identification Protocol

Previously reported protocols were applied (5 (link), 6 (link)). To identify podocytes, we used a combination of WT1 (Agilent Technologies; IS05530-2) and DACH1 (Sigma-Aldrich; HPA012672) (32 (link)) as primary antibodies; Alexa-Fluor 488, -555, and/or -647 as secondary antibodies (Invitrogen; A21202, A31572, A31571, and A31573, respectively) depending on the experiment; and a DNA marker to identify single nuclei — either DAPI (Sigma-Aldrich; D9542) or DRAQ5 (Abcam; ab108410). Optical images were obtained using inverted laser confocal microscopes (Nikon and LSM800, Zeiss), stored in 1024 × 1024 pixel frames. Each image contained 1 glomerulus.

Free full text: Click here

Zimmermann M., Klaus M., Wong M.N., Thebille A.K., Gernhold L., Kuppe C., Halder M., Kranz J., Wanner N., Braun F., Wulf S., Wiech T., Panzer U., Krebs C.F., Hoxha E., Kramann R., Huber T.B., Bonn S, & Puelles V.G. (2021). Deep learning–based molecular morphometrics for kidney biopsies. JCI Insight, 6(7), e144779.

Publication 2021

HPA012672 A21202 A31572 A31571 A31573 D9542 ab108410 LSM800

Alexa fluor 488 Antibodies Dapi Dna marker Frames Glomerulus Laser microscopes Podocytes Single nuclei

Corresponding Organization : Universität Hamburg

Other organizations : RWTH Aachen University, Martin Luther University Halle-Wittenberg, Sankt-Antonius-Hospital Eschweiler, Erasmus MC

Top 5 similar protocols

Protocol cited in 2 other protocols

Variable analysis

independent variables

Antibodies used to identify podocytes: WT1 and DACH1

dependent variables

Optical images of glomeruli

control variables

Laser confocal microscopes used to obtain images
Image resolution of 1024 × 1024 pixels
Each image containing 1 glomerulus

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!