Brain sections were stained with anti-pS129 α-syn as described above, with the exception that goat anti-rabbit secondary antibody conjugated with Alexa 647 (Thermo Fisher Scientific) was used (1:200 in blocking buffer) to avoid interference with LCO emission. PD sections were also stained with anti-α-syn antibody (Syn303, 1:1000, Biolegend), anti-phosphorylated tau antibody (AT100, 1:250, Thermo Fisher Scientific), anti-p62 antibody (1:100, BD Bioscience) and anti-TDP-43 antibody (1:500, Proteintech Group Inc.). After washing 3 × 5 min in PBS, they were incubated with 3 μM HS-68 for 30 min at room temperature. They were then washed with PBS and mounted with Dako mounting medium. Fluorescence images and emission spectra were collected using an inverted Zeiss LSM 780 confocal microscope.
Anti p62 antibody
Manufactured by BD
The Anti-p62 antibody is a laboratory reagent used in research applications. It targets the p62 protein, also known as sequestosome-1 (SQSTM1), which is involved in various cellular processes. This antibody can be used to detect and study the p62 protein in biological samples, but its specific applications and intended use should be determined by the researcher.
Automatically generated - may contain errors
3 protocols using anti p62 antibody
1
Multimodal Analysis of Synucleinopathy
2
Immunohistochemical Analysis of p62 and LC3 in Mouse Liver
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Immunohistochemistry of p62 and LC3 was performed on 5‐μm thick paraffin sections. The paraffin sections were preheated in a microwave oven for 10 minutes and were blocked with bovine serum albumin (1:200). Afterwards, the sections were incubated with anti‐p62 antibody (1:1000; BD Biosciences) and anti‐LC3 (1:1000; Novus) for the mouse liver sections. After primary antibody staining, peroxidase‐labelled polymer were used for signal detection and the sections were subsequently counterstained with haematoxylin for the nuclei. For semi‐quantitative analysis of p62 and LC3 accumulation, the scores were rated as grades 0 (none), 1 (minor), 2 (moderate) and 3 (severe). More than 10 sections in each mouse were evaluated. The scores were rated by two pathologists.
3
Comprehensive Brain Examination Protocol for bvFTD
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For one p.R406W patient with post-mortem brain examination, the protocol started with an 8–16-week fixation period using 10% buffered formalin. Five-micrometre slices were cut and a macroscopic quantification of localized atrophy took place. Samples from the frontal cortex, the temporal neocortex (superior temporal gyrus), the hippocampus, the striate area, the neostriatum, the basal ganglia, the substantia nigra, the thalamus, the mesencephalon, the pons, the medulla oblongata, the cerebellum and the spinal cord were studied. The latter region is not always studied as part of the standard protocol,39 (link),40 (link) but it was done in this case as the phenotype was bvFTD, which has been associated with concomitant motor neuron disease.
Immunohistochemical staining was performed to determine the nature of possible neuronal inclusions. To that end, anti-ubiquitin antibody (Dako), AT8 (for P-tau; Fujirebio Europe), 4G8 (for amyloid β; Signet), anti-FUS antibody (Proteintech Group Inc.), anti-trans-active response DNA binding protein of 43 kDa antibody (Proteintech Group Inc.) and anti-p62 antibody (BD Diagnostics) were used. The anti-FUS antibody is not always used as part of the standard protocol,39 (link),40 (link) but it is often included in bvFTD cases such as this one.
Immunohistochemical staining was performed to determine the nature of possible neuronal inclusions. To that end, anti-ubiquitin antibody (Dako), AT8 (for P-tau; Fujirebio Europe), 4G8 (for amyloid β; Signet), anti-FUS antibody (Proteintech Group Inc.), anti-trans-active response DNA binding protein of 43 kDa antibody (Proteintech Group Inc.) and anti-p62 antibody (BD Diagnostics) were used. The anti-FUS antibody is not always used as part of the standard protocol,39 (link),40 (link) but it is often included in bvFTD cases such as this one.
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