mab sha31  (Alpha Innotech)

Journal: International Journal of Molecular Sciences

Article Title: Cerebrospinal Fluid and Plasma Small Extracellular Vesicles and miRNAs as Biomarkers for Prion Diseases

doi: 10.3390/ijms22136822

Figure Lengend Snippet: Detection of the pathologic prion protein (PrP Sc /PrP res ) by Dot blot ( A ) and Western blot ( B ) in PMCA reactions seeded with serially diluted (1/5, 1/10 and 1/50) CSF-derived sEVs obtained from scrapie-affected sheep. Immunodetection was performed using the monoclonal Sha31 antibody. ( A ) Representative images of PrP res detection by Dot blot after four PMCA rounds of CSF-derived sEVs from one sheep at clinical stage of scrapie, one sheep at terminal stage, one sheep at preclinical stage and one negative sheep. A brain homogenate from scrapie sheep (10 −3 to 10 −9 diluted) is also shown for comparison. Some dots were subjected to Western blot for PrP res profile detection. ( B ) Representative images of PrP res detection by Western blot after four PMCA rounds of CSF-derived sEVs from one sheep at clinical stage of scrapie (1/10 dilution), one sheep at terminal stage (1/10 dilution), one sheep at preclinical stage (1/50 dilution), one negative sheep (1/10 dilution) and a brain homogenate from scrapie sheep (10 −6 and 10 −7 dilution). A proteinase K-digested classical scrapie isolate (Dawson strain) was used as positive control.

Article Snippet: PrP Sc detection was performed using Sha31 mAb conjugated to horseradish peroxidase (0.06 μg/mL), and ECL substrate (Pierce) was used to reveal peroxidase activity.

Techniques: Dot Blot, Western Blot, Derivative Assay, Immunodetection, Comparison, Positive Control

Journal: PLoS Pathogens

Article Title: Transgenic Rabbits Expressing Ovine PrP Are Susceptible to Scrapie

doi: 10.1371/journal.ppat.1005077

Figure Lengend Snippet: Fragments identified by mass spectrometry in the brains of scrapie-diseased tgOv rabbit and specific of the sheep sequence are colored. Bold: Sha31 anti-PrP epitope.

Article Snippet: Immunoblots revealed with the mAb Sha31 were quantified using the Alpha-Ease software (Alpha-Innotech, USA).

Techniques: Mass Spectrometry, Sequencing

Journal: PLoS Pathogens

Article Title: Transgenic Rabbits Expressing Ovine PrP Are Susceptible to Scrapie

doi: 10.1371/journal.ppat.1005077

Figure Lengend Snippet: (a) PrP C electrophoretic pattern and level of expression in the brain of wild-type, sheep (VRQ allele), tgOv rabbit and tg338 mice. The amounts of material loaded are indicated. (b) Two-dimensional electrophoretic gel analysis of PrP C from wild-type and tgOv rabbit. The equivalent of 1mg of brain extract was used for comparison (Acidic side at left). Blots were probed with Sha31 anti-PrP antibody.

Article Snippet: Immunoblots revealed with the mAb Sha31 were quantified using the Alpha-Ease software (Alpha-Innotech, USA).

Techniques: Expressing, Comparison

Journal: PLoS Pathogens

Article Title: Transgenic Rabbits Expressing Ovine PrP Are Susceptible to Scrapie

doi: 10.1371/journal.ppat.1005077

Figure Lengend Snippet: Midbrain sections from tgOv (a-b, e-f) and wild-type (c, g-i) rabbits challenged with LA21K fast scrapie prions and from a mock-infected tgOv rabbit (d, i-j). (a-d) PET blot analyses using monoclonal antibody Sha31 showed PrP res accumulation solely in LA21K fast challenged tgOv rabbits. (e-j) Hematoxylin and eosin-stained section at the level of the thalamus (e, g, i) and hippocampus (f, h, j) showing vacuolation, predominantly in the thalamus (e) and to a lesser extend in the hippocampus (f) of the LA21K fast challenged tgOv rabbits, but none in the control animals (g-j). Scale bar: 100 μm.

Article Snippet: Immunoblots revealed with the mAb Sha31 were quantified using the Alpha-Ease software (Alpha-Innotech, USA).

Techniques: Infection, Staining, Control

Journal: Journal of Virology

Article Title: Prion Propagation and Toxicity Occur In Vitro with Two-Phase Kinetics Specific to Strain and Neuronal Type

doi: 10.1128/JVI.03082-12

Figure Lengend Snippet: Monitoring of PrP aggregates in neurons and astrocytes in CGNC57BL/6 cultures infected with the 139A, ME7, or 22L scrapie strain. (A) Cultures were exposed to 139A-, ME7-, or 22L-infected brain homogenates at a final concentration of 0.01% (wt/vol). At different times postexposure, cells were fixed, permeabilized, and treated with 3 M guanidine thiocyanate. Immunostaining of PrP (in green) was performed by using monoclonal antibody Sha31. DAPI staining appears in blue. CGNC57BL/6 cultures infected with the 139A, ME7, and 22L strains are compared to control cultures (CT). Control and infected CGNC57BL/6 cultures shown in the figure were labeled at 21 dpe. In infected cultures, cells showed punctate fluorescence signals, suggesting intracellular PrP aggregates. In contrast, control cultures showed diffuse, homogenous PrP labeling. (B) Data presented in the box plots show the distribution of the quantified number of intraneuronal PrP aggregates at different days postexposure for each scrapie strain. The number of PrP aggregates per neuron significantly increased during the time course of infection in CGNC57BL/6 cells infected with the 139A, ME7, and 22L strains. In CGNC57BL/6 cells infected with the 22L strain, the variation of the number of PrP aggregates per neuron occurred earlier, as soon as 7 dpe, than with the other strains. The variations observed in ME7-infected CGNC57BL/6 cells were slighter than those observed with the two other strains. (C) Data represented in the box plot show the distribution of the size of intraneuronal PrP aggregates at different days after exposure to each scrapie strain. Regardless of the strain, infected CGNC57BL/6 cells showed a significant decrease in the size of neuronal PrP aggregates. (D) Data presented in the box plots show the distribution of the number of intra-astrocyte PrP aggregates at different days postexposure for each scrapie strain. Regardless of the strain, the number of PrP aggregates per astrocyte did not vary during the time course of infection. Box plots represent data from three independent experiments performed in triplicate ± the standard error of the mean. Differences compared between each point (*) were significant at a P value of ≤0.05.

Article Snippet: PrP was detected with MAb Sha31 (1/1,000; SpiBio, Massy, France).

Techniques: Infection, Concentration Assay, Immunostaining, Staining, Labeling, Fluorescence

Journal: Journal of Virology

Article Title: Prion Propagation and Toxicity Occur In Vitro with Two-Phase Kinetics Specific to Strain and Neuronal Type

doi: 10.1128/JVI.03082-12

Figure Lengend Snippet: Double immunofluorescence of PrP and organelle markers in CGNC57BL/6 cultures infected with the 139A strain at 21 dpe. Immunolabeled PrP (A, D, G, J, and M) and organelle markers (B, E, H, K, and N) were visualized by confocal microscopy (green and red, respectively). In merged images (C, F, I, L, and O), yellow areas indicate colocalization, and DAPI staining appears in blue. Immunostaining was performed with the following antibodies: Sha31 monoclonal antibody (PrP), giantin monoclonal antibody (Golgi apparatus) (B and C), early endosome autoantigen 1 monoclonal antibody (EEA1) (E and F), lysosome-associated membrane protein 2 monoclonal antibody (LAMP2) (H and I), cathepsin D monoclonal antibody (lysosomal lumen) (K and L), and GRP78 monoclonal antibody (endoplasmic reticulum) (N and O).

Article Snippet: PrP was detected with MAb Sha31 (1/1,000; SpiBio, Massy, France).

Techniques: Immunofluorescence, Infection, Immunolabeling, Confocal Microscopy, Staining, Immunostaining

Journal: Journal of Virology

Article Title: Prion Propagation and Toxicity Occur In Vitro with Two-Phase Kinetics Specific to Strain and Neuronal Type

doi: 10.1128/JVI.03082-12

Figure Lengend Snippet: Double immunofluorescence of PrP and organelle markers in CGNC57BL/6 cultures infected with the ME7 strain at 21 dpe. Immunolabeled PrP (A, D, G, J, and M) and organelle markers (B, E, H, K, and N) were visualized by confocal microscopy (green and red, respectively). In merged images (C, F, I, L, and O), yellow areas indicate colocalization, and DAPI staining appears in blue. Immunostaining was performed with the following antibodies: Sha31 monoclonal antibody (PrP), giantin monoclonal antibody (Golgi apparatus) (B and C), early endosome autoantigen 1 monoclonal antibody (EEA1) (E and F), lysosome-associated membrane protein 2 monoclonal antibody (LAMP2) (H and I), cathepsin D monoclonal antibody (lysosomal lumen) (K and L), and GRP78 monoclonal antibody (endoplasmic reticulum) (N and O).

Article Snippet: PrP was detected with MAb Sha31 (1/1,000; SpiBio, Massy, France).

Techniques: Immunofluorescence, Infection, Immunolabeling, Confocal Microscopy, Staining, Immunostaining

Journal: Journal of Virology

Article Title: Prion Propagation and Toxicity Occur In Vitro with Two-Phase Kinetics Specific to Strain and Neuronal Type

doi: 10.1128/JVI.03082-12

Figure Lengend Snippet: Double immunofluorescence of PrP and organelle markers in CGNC57BL/6 cultures infected with the 22L strain at 21 dpe. Immunolabeled PrP (A, D, G, J, and M) and organelle markers (B, E, H, K, and N) were visualized by confocal microscopy (green and red, respectively). In merged images (C, F, I, L, and O), yellow areas indicate colocalization, and DAPI staining appears in blue. Immunostaining was performed with the following antibodies: Sha31 monoclonal antibody (PrP), giantin monoclonal antibody (Golgi apparatus) (B and C), early endosome autoantigen 1 monoclonal antibody (EEA1) (E and F), lysosome-associated membrane protein 2 monoclonal antibody (LAMP2) (H and I), cathepsin D monoclonal antibody (lysosomal lumen) (K and L), and GRP78 monoclonal antibody (endoplasmic reticulum) (N and O).

Article Snippet: PrP was detected with MAb Sha31 (1/1,000; SpiBio, Massy, France).

Techniques: Immunofluorescence, Infection, Immunolabeling, Confocal Microscopy, Staining, Immunostaining