Immunohistochemical Analysis of DHHC21 and α1AR

Human renal arteries were fixed in 10% formalin for 48 h, paraffin-embedded and sectioned. Slides were then deparaffinized, rehydrated and permeabilized with PBS containing 0.05% Triton X-100^{22 (link)}. After blocking, slides were labelled with rabbit anti-DHHC21 and goat anti-α1AR antibodies (1:100) overnight at 4 °C. After washing, secondary antibody incubation was done with donkey anti-rabbit Alexa Fluor 488 and donkey anti-goat Alexa Fluor 568 (1:500). After mounting with ProLong Diamond mounting medium with DAPI, slides were imaged with Leica SP8 spectral inverted laser scanning confocal microscope.

Free full text: Click here

Yang X., Zheng E., Ma Y., Chatterjee V., Villalba N., Breslin J.W., Liu R., Wu M.H, & Yuan S.Y. (2021). DHHC21 deficiency attenuates renal dysfunction during septic injury. Scientific Reports, 11, 11146.

Publication 2021

SP8 spectral inverted laser scanning confocal microscope

Alexa 568 Alexa fluor 488 Anti antibodies Antibody Confocal laser scanning microscope Dapi Diamond Donkey Formalin Goat Human Paraffin Rabbit Renal arteries Slides

Corresponding Organization :

Other organizations : University of South Florida

Top 5 similar protocols

Variable analysis

independent variables

Tissue fixation in 10% formalin for 48 h
Paraffin embedding and sectioning of the tissue

dependent variables

Localization and expression of DHHC21 and α1AR proteins in the renal artery tissue

control variables

PBS containing 0.05% Triton X-100 for permeabilization
Blocking step before antibody labeling
Incubation of primary antibodies (rabbit anti-DHHC21 and goat anti-α1AR) overnight at 4°C
Incubation of secondary antibodies (donkey anti-rabbit Alexa Fluor 488 and donkey anti-goat Alexa Fluor 568) at 1:500 dilution
Mounting with ProLong Diamond mounting medium with DAPI
Imaging with Leica SP8 spectral inverted laser scanning confocal microscope

controls

Positive control: Not specified
Negative control: Not specified

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!