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.
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.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti smad2 3, by Cell Signaling Technology (1 mentions)
Fitc conjugated anti rabbit igg, by Merck Group (1 mentions)
Rabbit anti phospho smad3, by Cell Signaling Technology (1 mentions)
Rabbit anti smad1 5 8, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti mouse phospho stat1, by Cell Signaling Technology (1 mentions)
Facscanto 2, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Anti rabbit hrp conjugated antibody, by Cytiva (1 mentions)
4 protocols using rabbit anti mouse actin
1
Western Blot Analysis of Neutrophil LC3 and Actin
2
Multiparametric Analysis of Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
3
Western Blot Analysis of HIF-1α and Actin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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)].
4
Western Blot Analysis of Neutrophil LC3 and Actin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!