Thirty to 50 µg of protein (BCA assay; Pierce; Rockford, IL) was separated by 8% or 12% SDS-PAGE under reducing conditions and electrophoretically transferred to a PVDF membrane (Bio-Rad; Hercules, CA). Membranes were blocked (Odyssey® blocking buffer; at 25°C for 1 hr) and then probed (4°C overnight) with an anti-phospho-p38 MAPK rabbit polyclonal antibody (1:500 dilution; Cell Signaling) or an anti- NFκB p65 rabbit polyclonal antibody (1:200 dilution;Santa Cruz) and a mouse monoclonal antibody directed against β-actin (1:4000 dilution; Sigma; overnight). Species-specific secondary antibodies labeled with spectrally distinct IRDye® fluorescent dyes (LI-COR Biosciences; Lincoln, NE) were used to detect primary antibodies (1 hr at 25°C). Results were recorded on LI-COR ODYSSEY® Fc Imaging system (LI-COR Biosciences) and protein levels quantified using Image Studio 3.1(LI-COR Biosciences; Lincoln, NE). p-p38 MAPK and p65 levels were normalized to their respective β-actin levels. Antibodies directed against GAPDH were used to check for cytosolic protein contamination within the nuclear fraction and none was found (data not shown).
Anti phospho p38 mapk rabbit polyclonal antibody
Manufactured by Cell Signaling Technology
The Anti-phospho-p38 MAPK rabbit polyclonal antibody is a laboratory reagent used for the detection and analysis of phosphorylated p38 mitogen-activated protein kinase (MAPK) in biological samples. This antibody specifically recognizes the phosphorylated form of p38 MAPK, which is a key signaling molecule involved in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey fc imaging system, by LI COR (1 mentions)
Irdye fluorescent dyes, by LI COR (1 mentions)
Image studio 3, by LI COR (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Odyssey imaging system, by LI COR (1 mentions)
Mouse monoclonal anti α tubulin antibody, by Merck Group (1 mentions)
Irdye800 conjugated anti rabbit igg, by Rockland Immunochemicals (1 mentions)
Anti phospho creb, by Cell Signaling Technology (1 mentions)
2 protocols using anti phospho p38 mapk rabbit polyclonal antibody
1
Western Blot Analysis of Phosphorylated p38 MAPK and NF-kB p65
2
Quantifying Signaling Pathway Activation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lysates were solubilized in Laemmli sample buffer. Equal amounts of total protein, determined by a bicinchoninic acid (BCA) protein assay (Pierce, Rockford, IL) with BSA as the standard, were separated by SDS‐PAGE and electrotransfer of proteins from the gel to PVDF membranes. Blots were probed with anti‐p38 rabbit polyclonal MAPK (Cell Signaling, Hartsfordshire, U.K.), and anti‐phospho‐CREB (Cell Signaling) antibodies. Binding was detected with IRDye 800‐conjugated anti‐rabbit IgG (Rockland, Gilbertsville, PA) or IRDye 680‐conjugated anti‐mouse IgG (Invitrogen, Carlsbad, CA) secondary antibodies. All data are expressed as integrated intensity following infrared detection (Odyssey Imaging system; LI‐COR Biosciences, Lincoln, NE). For p38 MAPK signaling, membranes were then stripped (2% SDS (w/v) in 25 mmol/L glycine, pH 2.0) and reprobed with anti‐phospho‐p38 MAPK rabbit polyclonal antibody (Cell Signaling). As a loading control, blots were then reprobed with anti‐α‐tubulin mouse monoclonal antibody, which was not significantly different between groups (Sigma, Castle Hill, NSW, Australia).
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!