Samples were lysed in M-PER protein extraction buffer (Thermo Fisher Scientific). Proteins were separated on 12% polyacrylamide gels with Tris/SDS running buffer and transferred onto nitrocellulose (Invitrogen). Membranes were blocked with 5% milk, 3% bovine serum albumin (BSA) and 0.1% Tween 20 and incubated with polyclonal rabbit antimouse COL1A1 (91144S; Cell Signaling; diluted 1:1'000), monoclonal rabbit antimouse VIM (5741S, Cell Signaling; diluted 1:1'000), polyclonal rabbit antimouse TGFβ1 (3711S, Cell Signaling; diluted 1:1'000), polyclonal goat antimouse. IL-1β (AF-401-NA, Bio-techne, diluted 1:1'000) and polyclonal goat antihuman ACTA2 (NB300-978SS, Novus Biologicals, diluted 1:1'000). Horseradish peroxidase-conjugated secondary antibodies were donkey antigoat (#sc-2020; Santa Cruz; diluted 1:3'000) and goat antirabbit (#sc-2004; Santa Cruz; diluted 1:3'000). Additionally, β-Actin was used as a loading control.
Af 401 na
Manufactured by Bio-Techne
AF-401-NA is a laboratory instrument manufactured by Bio-Techne. It is designed for the detection and quantification of specific proteins or analytes in a sample. The core function of this product is to provide reliable and accurate measurement capabilities for researchers and scientists in various fields.
Automatically generated - may contain errors
2 protocols using af 401 na
1
Protein Extraction and Western Blot
2
Protein Quantification and Western Blotting
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Tissue was lysed in M-PER cell lysis buffer (Thermo Fisher Scientific).
Protein levels were determined by NanoDrop according to the manufacturer's instructions and equal amounts of protein were loaded onto SDS/PAGE gels.
Proteins were separated on 12% polyacrylamide gels with Tris/SDS running buffer and transferred onto nitrocellulose (Invitrogen). Membranes were blocked with 5% milk, 3% BSA and 0.1% Tween 20. WBs were performed using polyclonal rabbit anti-mouse IL6 (ab6627, Lucerna, 1:1000), polyclonal goat anti-mouse IL1β (AF-401-NA, R&D / bio-techne, 1:1000), polyclonal rabbit antimouse TGF-β (D157926 3711S BioConcept / CellSignaling, 1:1000), polyclonal
Protein levels were determined by NanoDrop according to the manufacturer's instructions and equal amounts of protein were loaded onto SDS/PAGE gels.
Proteins were separated on 12% polyacrylamide gels with Tris/SDS running buffer and transferred onto nitrocellulose (Invitrogen). Membranes were blocked with 5% milk, 3% BSA and 0.1% Tween 20. WBs were performed using polyclonal rabbit anti-mouse IL6 (ab6627, Lucerna, 1:1000), polyclonal goat anti-mouse IL1β (AF-401-NA, R&D / bio-techne, 1:1000), polyclonal rabbit antimouse TGF-β (D157926 3711S BioConcept / CellSignaling, 1:1000), polyclonal
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