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Ferritin is the most abundant non-PrP Sc protein found in all of the samples. A) The amount of PrP Sc relative to ferritin and other proteins was estimated by spectral counting and was found to vary between samples. Lower relative abundances of PrP Sc were associated with higher abundances of ferritin. In the case of CH-04, the amounts of ferritin, versican, and proteoglycan link proteins were much higher than in any of the other PrP Sc samples. An immunoblot from representative mock and PrP Sc samples were separated by SDS-PAGE on a 4–12% NuPAGE gradient gel and probed with B) anti-ferritin rabbit antibody, washed with <t>TBST</t> buffer and then probed with C) <t>6D11</t> anti-PrP mouse monoclonal antibody.
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Article Title: COMPARATIVE PROFILING OF HIGHLY ENRICHED 22L AND CHANDLER MOUSE SCRAPIE PRION PROTEIN PREPARATIONS

Journal: Proteomics

doi: 10.1002/pmic.201000104

Ferritin is the most abundant non-PrP Sc protein found in all of the samples. A) The amount of PrP Sc relative to ferritin and other proteins was estimated by spectral counting and was found to vary between samples. Lower relative abundances of PrP Sc were associated with higher abundances of ferritin. In the case of CH-04, the amounts of ferritin, versican, and proteoglycan link proteins were much higher than in any of the other PrP Sc samples. An immunoblot from representative mock and PrP Sc samples were separated by SDS-PAGE on a 4–12% NuPAGE gradient gel and probed with B) anti-ferritin rabbit antibody, washed with TBST buffer and then probed with C) 6D11 anti-PrP mouse monoclonal antibody.
Figure Legend Snippet: Ferritin is the most abundant non-PrP Sc protein found in all of the samples. A) The amount of PrP Sc relative to ferritin and other proteins was estimated by spectral counting and was found to vary between samples. Lower relative abundances of PrP Sc were associated with higher abundances of ferritin. In the case of CH-04, the amounts of ferritin, versican, and proteoglycan link proteins were much higher than in any of the other PrP Sc samples. An immunoblot from representative mock and PrP Sc samples were separated by SDS-PAGE on a 4–12% NuPAGE gradient gel and probed with B) anti-ferritin rabbit antibody, washed with TBST buffer and then probed with C) 6D11 anti-PrP mouse monoclonal antibody.

Techniques Used: SDS Page

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Western Blot:

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Article Snippet: .. Western blot membranes were blocked with 5% BSA in TBST for at least 1 h and were extensively wash (four times) with TBST 10 min prior to detection with Clarity Western ECL substrates (Bio-Rad Laboratories, Hercules, California, USA). .. ALPHAScreen ALPHAScreen was performed as previously described – , using the cMyc detection kit and Proxiplate-384 Plus plates (PerkinElmer).

Article Title: COMPARATIVE PROFILING OF HIGHLY ENRICHED 22L AND CHANDLER MOUSE SCRAPIE PRION PROTEIN PREPARATIONS
Article Snippet: .. The PVDF membrane used for ferritin western blot was then washed in TBST, and re-probed with anti-PrP monoclonal antibody 6D11 at 1:10,000, anti-mouse IgG at 1:200,000, and Streptactin-HRP (BIO-RAD) at 1:400,000. .. Western C molecular weight standards were used to align scanned images of the ferritin and PrP stains.

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Incubation:

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Imaging:

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    Bio-Rad tbst
    Ferritin is the most abundant non-PrP Sc protein found in all of the samples. A) The amount of PrP Sc relative to ferritin and other proteins was estimated by spectral counting and was found to vary between samples. Lower relative abundances of PrP Sc were associated with higher abundances of ferritin. In the case of CH-04, the amounts of ferritin, versican, and proteoglycan link proteins were much higher than in any of the other PrP Sc samples. An immunoblot from representative mock and PrP Sc samples were separated by SDS-PAGE on a 4–12% NuPAGE gradient gel and probed with B) anti-ferritin rabbit antibody, washed with <t>TBST</t> buffer and then probed with C) <t>6D11</t> anti-PrP mouse monoclonal antibody.
    Tbst, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1311 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tbst/product/Bio-Rad
    Average 99 stars, based on 1311 article reviews
    Price from $9.99 to $1999.99
    tbst - by Bioz Stars, 2020-05
    99/100 stars
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    Ferritin is the most abundant non-PrP Sc protein found in all of the samples. A) The amount of PrP Sc relative to ferritin and other proteins was estimated by spectral counting and was found to vary between samples. Lower relative abundances of PrP Sc were associated with higher abundances of ferritin. In the case of CH-04, the amounts of ferritin, versican, and proteoglycan link proteins were much higher than in any of the other PrP Sc samples. An immunoblot from representative mock and PrP Sc samples were separated by SDS-PAGE on a 4–12% NuPAGE gradient gel and probed with B) anti-ferritin rabbit antibody, washed with TBST buffer and then probed with C) 6D11 anti-PrP mouse monoclonal antibody.

    Journal: Proteomics

    Article Title: COMPARATIVE PROFILING OF HIGHLY ENRICHED 22L AND CHANDLER MOUSE SCRAPIE PRION PROTEIN PREPARATIONS

    doi: 10.1002/pmic.201000104

    Figure Lengend Snippet: Ferritin is the most abundant non-PrP Sc protein found in all of the samples. A) The amount of PrP Sc relative to ferritin and other proteins was estimated by spectral counting and was found to vary between samples. Lower relative abundances of PrP Sc were associated with higher abundances of ferritin. In the case of CH-04, the amounts of ferritin, versican, and proteoglycan link proteins were much higher than in any of the other PrP Sc samples. An immunoblot from representative mock and PrP Sc samples were separated by SDS-PAGE on a 4–12% NuPAGE gradient gel and probed with B) anti-ferritin rabbit antibody, washed with TBST buffer and then probed with C) 6D11 anti-PrP mouse monoclonal antibody.

    Article Snippet: The PVDF membrane used for ferritin western blot was then washed in TBST, and re-probed with anti-PrP monoclonal antibody 6D11 at 1:10,000, anti-mouse IgG at 1:200,000, and Streptactin-HRP (BIO-RAD) at 1:400,000.

    Techniques: SDS Page