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Rabbit anti mouse actin

Manufactured by Merck Group
Sourced in Germany

Rabbit anti-mouse actin is a primary antibody that recognizes the actin protein found in mouse cells. It is commonly used in research applications for the detection and quantification of actin in various experimental systems.

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4 protocols using rabbit anti mouse actin

1

Western Blot Analysis of Neutrophil LC3 and Actin

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Whole cell lysates from isolated neutrophils were run on a 4-12% Bis-Tris gel and transferred to a 0.2 mm nitrocellulose membrane. After blocking with 5% milk, membranes were incubated overnight at 4° C with primary antibodies specific for rabbit anti-mouse LC3 (Sigma, #L8918) and rabbit anti-mouse actin (Sigma, #A2066). Membranes were then incubated with peroxidase-conjugated secondary antibodies for 1 hour at 25° C and developed with the SuperSignal West Pico chemiluminescence kit (Pierce, #34079) and exposed on film.
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2

Multiparametric Analysis of Signaling Pathways

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Rabbit anti phospho-Smad2 (Millipore, Billerica, MA, USA), rabbit anti Smad2/3 (Cell Signaling Technology, Danvers, MA, USA), rabbit anti phospho-Smad3 (Cell Signaling Technology), goat anti phospho-Smad1/5/8 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti Smad1/5/8 (Santa Cruz Biotechnology), rabbit anti-mouse phospho-Stat1 (Cell Signaling Technology), rabbit anti-mouse ApoA-I (Santa Cruz Biotechnology), rabbit anti-mouse Actin (Sigma, Steinheim, Germany) and HRP conjugate donkey anti-rabbit IgG (Southern Biotech, Birmingham, AL, USA) were used for western blot analysis. Rabbit anti SRB1 monoclonal antibody (Novus Biologicals, CO, USA) and FITC conjugated anti-rabbit IgG (Sigma, Steinheim, Germany) were used for flow cytometry analysis. Fc block and fluorochrome conjugated antibodies against mouse CD45, CD3, CD8, MHC-II, CD11c and TNF-alpha were purchased from BD Biosciences. Intracellular staining for TNF-alpha was performed following manufacturer's instructions. For flow cytometry experiments acquisition was done using either FACSCalibur or FACSCanto II (BD Biosciences). FlowJo Software (TreeStar) was used to analyze cellular events.
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3

Western Blot Analysis of HIF-1α and Actin

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Cells were lysed in a buffer containing 140 mM NaCl, 20 mM Tris-HCl pH 7.4, 5 mM EDTA, 10% glycerol, 1% Triton X-100. Forty μg of cell lysate were loaded and subjected to SDS-PAGE and Western blotting (WB). The membranes were hybridized overnight with rabbit anti-mouse actin (1:5,000; Sigma-Aldrich) and rabbit anti-mouse HIF-1α (1:1,000; Novus Biologicals), followed by incubation with anti-rabbit HRP-conjugated antibody (1:5,000 dilution, Amersham-Pharmacia, Little Chalfont, U.K.). The signal was finally detected by chemiluminescence with SuperSignal kit (Pierce, Rockford, IL, USA) and lane densitometry analyzed by standard procedures [49 (link)].
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4

Western Blot Analysis of Neutrophil LC3 and Actin

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Whole cell lysates from isolated neutrophils were run on a 4-12% Bis-Tris gel and transferred to a 0.2 mm nitrocellulose membrane. After blocking with 5% milk, membranes were incubated overnight at 4° C with primary antibodies specific for rabbit anti-mouse LC3 (Sigma, #L8918) and rabbit anti-mouse actin (Sigma, #A2066). Membranes were then incubated with peroxidase-conjugated secondary antibodies for 1 hour at 25° C and developed with the SuperSignal West Pico chemiluminescence kit (Pierce, #34079) and exposed on film.
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