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Oneblock western fl blocking buffer

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OneBlock Western-FL Blocking Buffer is a pre-made solution designed to block non-specific protein binding in Western blot analysis. It is an effective blocking agent that helps to reduce background signal and improve the specificity of the antibody-antigen interaction.

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Lab products found in correlation

Chemidoc xrs imager, by Bio-Rad (4 mentions) Western ecl substrate, by Bio-Rad (4 mentions) Precast 4 20 polyacrylamide gels, by Bio-Rad (2 mentions) Anti hsp27, by Cell Signaling Technology (2 mentions) Anti o glcnac rl2, by Thermo Fisher Scientific (2 mentions) Anti αbc, by Cell Signaling Technology (2 mentions) Mabn640, by Merck Group (2 mentions) Hrp conjugated anti rabbit secondary antibody, by Jackson ImmunoResearch (2 mentions) Tbs t, by Cell Signaling Technology (2 mentions) Tbs t, by Bio-Rad (2 mentions)

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OneBlock blocking buffer (5 citations) Oneblock buffer (2 citations) OneBlock (2 citations)

4 protocols using oneblock western fl blocking buffer

1

Western Blotting of Brain Proteins

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Input and pulldown gel samples were bath sonicated, boiled for 10 min, and separated by SDS-PAGE on precast 4-20% polyacrylamide gels (BioRad). Proteins were then transferred on PVDF membranes using a semi-dry transfer apparatus. For dot-blotting, each spot was added with 5 ug of brain lysate protein on a nitrocellulose membrane and allowed to air-dry for 1.5 hours before blocking. Membranes were blocked for 1 h at room temperature using OneBlock Western-FL Blocking Buffer (Genesee Scientific) after which the membranes were incubated with primary antibodies (Anti-O-GlcNAc RL2 1:3,000, Thermo MA1-072; Anti-Hsp27 1:3,000, Cell Signaling Technology #95357; Anti-αBC 1:3,000, Cell Signaling Technology #45584) in blocking buffer at 4 °C for 16 h. Membranes were washed with TBST (137 mM NaCl, 20 mM Tris, 0.1% Tween-20, pH 7.6, Cell Signaling Technology) 3 times for 10 min each, followed by incubation with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (1:10,000, Jackson ImmunoResearch 711-035-152 or 715-035-150) in blocking buffer. After 3x10 min washing in TBST, membranes were developed with Western ECL Substrates (Biorad) and imaged using a ChemiDoc XRS+ Imager (Bio-Rad).
Balana A.T., Levine P.M., Craven T.W., Mukherjee S., Pedowitz N.J., Moon S.P., Takahashi T.T., Becker C.F., Baker D, & Pratt M.R. (2021). O-GlcNAc modification of small heat shock proteins enhances their anti-amyloid chaperone activity. Nature chemistry, 13(5), 441-450.
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2

Amyloid Beta Aggregate Analysis

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Blot samples were prepared by diluting the amyloid beta aggregation reaction at the time points indicated in Figure 4b four-fold into 4% SDS buffer (3% final SDS concentration). For each spot, a volume corresponding to 10 ng of amyloid beta (based on monomer concentration) was applied on a dry nitrocellulose membrane, before air-drying for 30 minutes. The membrane was then blocked with OneBlock Western-FL Blocking Buffer (Genesee Scientific) for 1 hour at room temperature. Anti-amyloid beta fibril-specific antibody (EMD Millipore, MABN640) was added at 1:10,000 dilution in OneBlock buffer and incubated for 16 hours at 4°C. The membrane was washed three times for 5 minutes each in TBST (Cell Signaling Technologies) and secondary anti-rabbit HRP-conjugated antibody (Jackson) was added at 1:10,000 dilution in OneBlock buffer. After 1 hour at room temperature, the membrane was washed three times for 5 minutes in TBST, developed with Western ECL Substrates (Biorad) and imaged using a ChemiDoc XRS+ Imager (Bio-Rad)
Balana A.T., Levine P.M., Craven T.W., Mukherjee S., Pedowitz N.J., Moon S.P., Takahashi T.T., Becker C.F., Baker D, & Pratt M.R. (2021). O-GlcNAc modification of small heat shock proteins enhances their anti-amyloid chaperone activity. Nature chemistry, 13(5), 441-450.
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3

Western Blotting of Brain Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Input and pulldown gel samples were bath sonicated, boiled for 10 min, and separated by SDS-PAGE on precast 4-20% polyacrylamide gels (BioRad). Proteins were then transferred on PVDF membranes using a semi-dry transfer apparatus. For dot-blotting, each spot was added with 5 ug of brain lysate protein on a nitrocellulose membrane and allowed to air-dry for 1.5 hours before blocking. Membranes were blocked for 1 h at room temperature using OneBlock Western-FL Blocking Buffer (Genesee Scientific) after which the membranes were incubated with primary antibodies (Anti-O-GlcNAc RL2 1:3,000, Thermo MA1-072; Anti-Hsp27 1:3,000, Cell Signaling Technology #95357; Anti-αBC 1:3,000, Cell Signaling Technology #45584) in blocking buffer at 4 °C for 16 h. Membranes were washed with TBST (137 mM NaCl, 20 mM Tris, 0.1% Tween-20, pH 7.6, Cell Signaling Technology) 3 times for 10 min each, followed by incubation with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (1:10,000, Jackson ImmunoResearch 711-035-152 or 715-035-150) in blocking buffer. After 3x10 min washing in TBST, membranes were developed with Western ECL Substrates (Biorad) and imaged using a ChemiDoc XRS+ Imager (Bio-Rad).
Balana A.T., Levine P.M., Craven T.W., Mukherjee S., Pedowitz N.J., Moon S.P., Takahashi T.T., Becker C.F., Baker D, & Pratt M.R. (2021). O-GlcNAc modification of small heat shock proteins enhances their anti-amyloid chaperone activity. Nature chemistry, 13(5), 441-450.
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4

Amyloid Beta Aggregate Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Blot samples were prepared by diluting the amyloid beta aggregation reaction at the time points indicated in Figure 4b four-fold into 4% SDS buffer (3% final SDS concentration). For each spot, a volume corresponding to 10 ng of amyloid beta (based on monomer concentration) was applied on a dry nitrocellulose membrane, before air-drying for 30 minutes. The membrane was then blocked with OneBlock Western-FL Blocking Buffer (Genesee Scientific) for 1 hour at room temperature. Anti-amyloid beta fibril-specific antibody (EMD Millipore, MABN640) was added at 1:10,000 dilution in OneBlock buffer and incubated for 16 hours at 4°C. The membrane was washed three times for 5 minutes each in TBST (Cell Signaling Technologies) and secondary anti-rabbit HRP-conjugated antibody (Jackson) was added at 1:10,000 dilution in OneBlock buffer. After 1 hour at room temperature, the membrane was washed three times for 5 minutes in TBST, developed with Western ECL Substrates (Biorad) and imaged using a ChemiDoc XRS+ Imager (Bio-Rad)
Balana A.T., Levine P.M., Craven T.W., Mukherjee S., Pedowitz N.J., Moon S.P., Takahashi T.T., Becker C.F., Baker D, & Pratt M.R. (2021). O-GlcNAc modification of small heat shock proteins enhances their anti-amyloid chaperone activity. Nature chemistry, 13(5), 441-450.
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