primary antibody sha31  (Bio-Rad)

Journal: Acta neuropathologica

Article Title: Characterization of prion strains and peripheral prion infectivity patterns in E200K genetic CJD patients.

doi: 10.1007/s00401-025-02903-5

Figure Lengend Snippet: Fig. 1 PrPres western blot profiles in original E200K gCJD isolates and E200K gCJD-inoc- ulated human PrP-expressing mice. Western blot analysis of proteinase K-resistant prion protein (PrPres) profiles was performed on brain homogen- ates from both the original E200K gCJD patient isolates and transgenic mice expressing human PrP with Met129 (tgMet) or Val129 (tgVal) at codon 129. Mice were intracerebrally inoculated with 20 µL of 10% brain homogenate from E200K gCJD patients (n = 6 per group). Two serial passages were carried out in each mouse line (details in Table 1). The PrPres isoform (type 1 or type 2) was identified via SDS-PAGE and Western blot using the anti-PrP monoclonal antibody Sha31 (epitope YEDRYYRE). To control for PrPres isoform, MM1 sCJD (type 1) and VV2 sCJD (type 2) isolates were included in each gel. The PrPres isoform results for each passage and mouse line are summarized in Table 1

Article Snippet: Immunodetection was carried on PVDF membranes out using the monoclonal primary antibody Sha31 (1 μg/mL), which recognizes amino acids 145–152 (YEDRYYRE) of PrP, and an anti-mouse HRP-conjugated secondary antibody (Biorad) [16].

Techniques: Western Blot, Expressing, Transgenic Assay, SDS Page, Control

Journal: Acta neuropathologica

Article Title: Characterization of prion strains and peripheral prion infectivity patterns in E200K genetic CJD patients.

doi: 10.1007/s00401-025-02903-5

Figure Lengend Snippet: Fig. 3 PrPres western blot in the brains of tgMet mice inoculated with brain and peripheral tissues from E200K gCJD Patients. Western blot analysis of proteinase K-resistant prion protein (PrPres) was conducted on brain homogenates from transgenic mice expressing human PrP with Met129 (tgMet) after two serial intracerebral passages (n = 6 per group). The mice were inoculated with 20µL of a 10% homogenate prepared from brain or peripheral tissues of E200K gCJD patients (refer to Tables 1 and 2). PrPres was detected using the anti-PrP mon- oclonal antibody Sha31, targeting the YEDRYYRE epitope. For com- parison, MM1 sCJD (type 1) and VV2 sCJD (type 2) prion isolates were included as controls. PrPres isoform identification results for each passage are summarized in Table 2

Article Snippet: Immunodetection was carried on PVDF membranes out using the monoclonal primary antibody Sha31 (1 μg/mL), which recognizes amino acids 145–152 (YEDRYYRE) of PrP, and an anti-mouse HRP-conjugated secondary antibody (Biorad) [16].

Techniques: Western Blot, Transgenic Assay, Expressing

Journal: Marine Drugs

Article Title: Potential of Marine Sponge Metabolites against Prions: Bromotyrosine Derivatives, a Family of Interest

doi: 10.3390/md22100456

Figure Lengend Snippet: Anti-PrP Sc activity of compounds 1 – 6 on prion-infected MovS6 cells. MovS6 cells were treated with the indicated ranges of concentrations of the molecules, and absolute ethanol was used as a negative control. After 6 days of culture, cell lysates were digested by proteinase K to reveal PrP Sc (top panels) or untreated to reveal PrP C (middle panels) or the loading control Tubulin (bottom panels). Proteins were separated in 10% Bis-Tris polyacrylamide gels and revealed using anti-PrP (Sha31) or anti-tubulin-specific antibodies. The blots shown are representative of two to three independent experiments that all produced similar results. Purealidin Q ( 1 ), aplysamine-2 ( 2 ) and aplysamine-1 ( 5 ) were able to reduce PrP Sc propagation, whereas pseudoceratinine A ( 3 ), aerophobin-2 ( 4 ), and pseudoceratinine-B ( 6 ) were not.

Article Snippet: Membranes were then incubated overnight at 4 °C with 1/40,000 Sha31 primary antibody (Bertin Pharma, Montigny le Bretonneux, France, #A03213) in TBS-T (10 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.05% Tween 20).

Techniques: Activity Assay, Infection, Negative Control, Control, Produced

Journal: Emerging Infectious Diseases

Article Title: Scrapie versus Chronic Wasting Disease in White-Tailed Deer

doi: 10.3201/eid3008.240007

Figure Lengend Snippet: Epitope mapping on Western blots with antibodies that progress toward the N-terminal of the prion protein in study of scrapie versus CWD in white-tailed deer. C-terminal antibodies 6H4 (A) and SHA31 (B) were used to probe brain and lymphoid tissue (representative samples). C) Material from the second passage of WTD scrapie and CWD, both responsive to probing by the N-terminal antibody 12B2. D) WTD scrapie material showing no or low affinity to the N-terminal antibody P4. Cbrum, cerebrum; CWD, chronic wasting disease; P1, first passage; rpln, retropharyngeal lymph node; sc, cervical spinal cord; WTD, white-tailed deer.

Article Snippet: After 4 PBS washes, we blocked membranes with 5% nonfat milk in in Tris-buffered saline with 0.05% TWEEN 20 for 1 hour, then probed at 4°C overnight with SHA31 (Bertin Technologies, https://www.bertin-technologies.fr ) diluted 1∶5,000, followed by horseradish peroxidase–conjugated goat antimouse IgG secondary antibody.

Techniques: Western Blot