Quadriceps muscles (~50 mg) were homogenized on ice, protein was extracted, and samples were prepared for western blotting as performed previously.9 Extracted proteins (30 μg) were electrophoresed, transferred onto nitrocellulose membranes, and prepared for antibody incubations as carried out previously.9 Antibodies used were PGC1α (#AB3242) from MilliporeSigma (Burlington, MA, USA); OPA1 (#80471), Mitofusin‐2 (#9482), VDAC (#4866), cytochrome‐C (#11940) and Cox IV (#4844) from Cell Signalling Technologies (Danvers, MA, USA); and α‐Tubulin (#12G10) from Developmental Studies Hybridoma Bank (Iowa City, IA, USA). Membranes were then incubated with either anti‐rabbit IgG (H + L) DyLight 800 or anti‐mouse IgG (H + L) DyLight 680 secondary antibodies (Cell Signalling Technologies, Danvers, MA, USA), and analysed with Odyssey's Infrared Imaging System (LI‐COR Biosciences, Lincoln, NE, USA). Total proteins were normalized to the tubulin loading control.
Anti mouse igg h l dylight 680
Manufactured by Cell Signaling Technology
Sourced in United States
Anti‐mouse IgG (H + L) DyLight 680 is a secondary antibody conjugated to the DyLight 680 fluorescent dye. It is designed to detect and visualize mouse primary antibodies in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti rabbit igg h l dylight 800, by Cell Signaling Technology (2 mentions)
Cytochrome c, by Cell Signaling Technology (1 mentions)
Cox 4, by Cell Signaling Technology (1 mentions)
α tubulin, by Developmental Studies Hybridoma Bank (1 mentions)
Mitofusin 2, by Cell Signaling Technology (1 mentions)
S odyssey infrared imaging system, by LI COR (1 mentions)
Pgc 1α, by Merck Group (1 mentions)
Integrin β1, by Cell Signaling Technology (1 mentions)
Odyssey clx imaging system, by LI COR (1 mentions)
Phospho smad2 ser465 467, by Cell Signaling Technology (1 mentions)
Tgfbr2, by Abcam (1 mentions)
Tgfbr1, by Santa Cruz Biotechnology (1 mentions)
Smad2, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Cell Signaling Technology (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Phosphatase inhibitor cocktail, by Roche (1 mentions)
2 protocols using anti mouse igg h l dylight 680
1
Quantifying Mitochondrial Proteins in Muscle
2
Comprehensive Protein Expression Analysis
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Cell lysates were prepared with RIPA buffer (Cell Signaling, #9806), supplemented with 1 mM PMSF, 1 mM NaF, protease inhibitor cocktail (Roche, #04 693 132 001), and phosphatase inhibitor cocktail (Roche, #04 906 845 001). SDS–PAGE gels were transferred onto nitrocellulose membranes.
We used the following primary antibodies against: Phospho-SMAD2 (Ser465/467) (Cell Signaling, #3108), SMAD2 (Cell Signaling, #3103), β-Actin (Cell Signaling, #3700), TGFBR1 (Santa Cruz, #sc-398), TGFBR2 (abcam, #ab184948), and integrin β1 (Cell Signaling, #9699). All Western blot membranes were incubated with primary antibodies overnight at 4 °C. Secondary antibodies used for Western blotting were anti-rabbit IgG (H+L) DyLight 800 (Cell Signaling, #5151) and anti-mouse IgG (H+L) DyLight 680 (Cell Signaling, #5470). Western blot images were acquired and quantified using Odyssey CLx Imaging System (LI-COR Biosciences, #9140).
We used the following primary antibodies against: Phospho-SMAD2 (Ser465/467) (Cell Signaling, #3108), SMAD2 (Cell Signaling, #3103), β-Actin (Cell Signaling, #3700), TGFBR1 (Santa Cruz, #sc-398), TGFBR2 (abcam, #ab184948), and integrin β1 (Cell Signaling, #9699). All Western blot membranes were incubated with primary antibodies overnight at 4 °C. Secondary antibodies used for Western blotting were anti-rabbit IgG (H+L) DyLight 800 (Cell Signaling, #5151) and anti-mouse IgG (H+L) DyLight 680 (Cell Signaling, #5470). Western blot images were acquired and quantified using Odyssey CLx Imaging System (LI-COR Biosciences, #9140).
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