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Western blot block buffer

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Western blot block buffer is a ready-to-use solution designed to block non-specific binding sites on a membrane during Western blot analysis. It is formulated to effectively reduce background signals and enhance the specificity of antibody detection.

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Block buffer (2 citations)

2 protocols using western blot block buffer

1

Protein Expression Analysis by Western Blot

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Prior to transfection, cells were plated at a density of 5 × 105 cells per well in 24-well plates or 1 × 106 per P-100 tissue culture dish and incubated overnight in complete RPMI media. Following overnight incubation, fresh media was added and transfection or infection was performed. Following treatment, the cells were lysed in lysis buffer containing 0.5% NP-40. Total cellular protein was determined using the bicinchoninic acid (BCA) protein assay (Pierce, Rockford IL) and 10 µg of total protein was resolved by electrophoresis on a 7.5% SDS-PAGE gel at 150 volts constant at 4o C. Proteins were transferred to nitrocellulose at 100 volts followed by blocking in Licor Western blot block buffer. Immunoblot analysis was also performed on the Licor Odyssey platform using monoclonal anti-MR antibodies and goat anti-mouse IR dyes (Lincoln NE).
Vigerust D.J., Vick S, & Shepherd V.L. (2015). Stable Expression and Characterization of an Optimized Mannose Receptor. Journal of clinical & cellular immunology, 6(3), 330.
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2

Protein Expression Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Prior to transfection, cells were plated at a density of 5 × 105 cells per well in 24-well plates or 1 × 106 per P-100 tissue culture dish and incubated overnight in complete RPMI media. Following overnight incubation, fresh media was added and transfection or infection was performed. Following treatment, the cells were lysed in lysis buffer containing 0.5% NP-40. Total cellular protein was determined using the bicinchoninic acid (BCA) protein assay (Pierce, Rockford IL) and 10 µg of total protein was resolved by electrophoresis on a 7.5% SDS-PAGE gel at 150 volts constant at 4o C. Proteins were transferred to nitrocellulose at 100 volts followed by blocking in Licor Western blot block buffer. Immunoblot analysis was also performed on the Licor Odyssey platform using monoclonal anti-MR antibodies and goat anti-mouse IR dyes (Lincoln NE).
Vigerust D.J., Vick S, & Shepherd V.L. (2015). Stable Expression and Characterization of an Optimized Mannose Receptor. Journal of clinical & cellular immunology, 6(3), 330.
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