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: Prion

Article Title: Cellular prion protein distribution in the vomeronasal organ, parotid, and scent glands of white-tailed deer and mule deer

doi: 10.1080/19336896.2022.2079888

Figure Lengend Snippet: Relative pixel intensity analysis of PrP C expression in facial integumentary clarified 10% (w/v) gland homogenates of mule deer (A, C, E) and white-tailed deer (B, D, F). Background-adjusted average pixel intensities of PrP C bands of forehead (a-b), preorbital (c-d), and vestibular nasal glands were compared between lanes of SDS-PAGE PVDF membranes probed with anti-PrP Sha31. Unclarified white-tailed deer whole brain homogenate was used for reference. Sample size, mean, 95% confidence intervals, and significance by Mann-Whitney tests are shown.

Article Snippet: PrP C was probed with the mouse IgG 1 anti-prion protein SHA31 monoclonal primary antibody (1:10,000) (Cayman Chemical, USA) which binds to an epitope of 148–155 ( Odocoileus amino acid sequence).

Techniques: Expressing, SDS Page, MANN-WHITNEY

Journal: Prion

Article Title: Cellular prion protein distribution in the vomeronasal organ, parotid, and scent glands of white-tailed deer and mule deer

doi: 10.1080/19336896.2022.2079888

Figure Lengend Snippet: Protein concentration-adjusted PrP C protein expression in deer exocrine glands. Capillary electrophoresis immunoassay chemiluminescence sample size, mean, and 95% confidence intervals of clarified 10% (w/v) deer gland homogenates prepared in RIPA buffer. Deer facial (a-e) and leg (f-h) tissue and gland homogenate samples were adjusted to final protein concentrations of 1.5 μg/μL for the immunoassay. PrP C signal was detected by anti-PrP SHA31 antibody.

Article Snippet: PrP C was probed with the mouse IgG 1 anti-prion protein SHA31 monoclonal primary antibody (1:10,000) (Cayman Chemical, USA) which binds to an epitope of 148–155 ( Odocoileus amino acid sequence).

Techniques: Protein Concentration, Expressing, Electrophoresis

Journal: Prion

Article Title: Cellular prion protein distribution in the vomeronasal organ, parotid, and scent glands of white-tailed deer and mule deer

doi: 10.1080/19336896.2022.2079888

Figure Lengend Snippet: Species and sex influence on PrP C detection by anti-PrP SHA31 capillary gel electrophoresis assay with total protein concentration-standardized 10% (w/v) clarified gland homogenates

Article Snippet: PrP C was probed with the mouse IgG 1 anti-prion protein SHA31 monoclonal primary antibody (1:10,000) (Cayman Chemical, USA) which binds to an epitope of 148–155 ( Odocoileus amino acid sequence).

Techniques: Nucleic Acid Electrophoresis, Concentration Assay

Journal: Prion

Article Title: Cellular prion protein distribution in the vomeronasal organ, parotid, and scent glands of white-tailed deer and mule deer

doi: 10.1080/19336896.2022.2079888

Figure Lengend Snippet: Quantified PrP C protein concentration in deer gland homogenates. Total PrP C concentrations of individuals, means, and 95% confidence intervals of clarified 10% (w/v) mule deer (MD) and white-tailed deer (WT) facial (a-e) and leg (f-h) gland homogenates prepared in RIPA buffer as determined by SHA31-N5 sandwich ELISA. Protein concentration was calculated using a full-length recombinant deer prion protein standard curve.

Article Snippet: PrP C was probed with the mouse IgG 1 anti-prion protein SHA31 monoclonal primary antibody (1:10,000) (Cayman Chemical, USA) which binds to an epitope of 148–155 ( Odocoileus amino acid sequence).

Techniques: Protein Concentration, Sandwich ELISA, Recombinant

Journal: Prion

Article Title: Cellular prion protein distribution in the vomeronasal organ, parotid, and scent glands of white-tailed deer and mule deer

doi: 10.1080/19336896.2022.2079888

Figure Lengend Snippet: Mean PrP C concentrations and 95% confidence intervals of clarified 10% (w/v) gland homogenates as determined by SHA31-N5 sandwich ELISA

Article Snippet: PrP C was probed with the mouse IgG 1 anti-prion protein SHA31 monoclonal primary antibody (1:10,000) (Cayman Chemical, USA) which binds to an epitope of 148–155 ( Odocoileus amino acid sequence).

Techniques: Concentration Assay