Protein concentrations were measured with a BCA assay kit (Beyotime, Shanghai China). Equal amounts of denatured protein were subjected to 10% SDS-PAGE and blotted onto PVDF membranes (Millipore, Bedford, MA, USA). Antibodies against ST6Gal-I (Santa Cruz Biotech, USA), BACE1, VE-Cadherin, phospho-PKCδ, PKCδ, phospho-MEK1/2, MEK1/2, phospho-ERK44/42, ERK44/42 (Cell Signaling Technology, USA) and β-actin (Beyotime, Shanghai, China) were used as primary antibodies, and horseradish peroxidase-conjugated goat α-rabbit IgG (Beyotime, Shanghai, China) was used as the secondary antibody. Detection was performed using an ECL kit (Millipore, USA), according to manufacturer’s instructions. The relative amount of protein was determined by densitometry using Labworks software.
Phospho erk44 42
Manufactured by Cell Signaling Technology
Sourced in United States
Phospho-ERK44/42 is a lab equipment product that detects the phosphorylation of ERK1/2 (Extracellular signal-Regulated Kinase 1/2) proteins. It is used to measure the activation of the ERK signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Phospho mek1 2, by Cell Signaling Technology (1 mentions)
Phospho pkcδ, by Cell Signaling Technology (1 mentions)
Erk 44 42, by Cell Signaling Technology (1 mentions)
Bace1, by Cell Signaling Technology (1 mentions)
Ecl kit, by Merck Group (1 mentions)
Ve cadherin, by Cell Signaling Technology (1 mentions)
Mek1 2, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
β actin, by Beyotime (1 mentions)
Bca assay kit, by Beyotime (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
Wortmannin, by Merck Group (1 mentions)
Phospho akt ser, by Cell Signaling Technology (1 mentions)
Supersignal west pico chemiluminescent, by Thermo Fisher Scientific (1 mentions)
Smad7, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Merck Group (1 mentions)
Foxo3a, by Cell Signaling Technology (1 mentions)
2 protocols using phospho erk44 42
1
Protein Expression Analysis by Western Blot
2
Western Blot Analysis of Cell Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein expression was investigated by western blot analysis using 10–40 μg of total protein. Tissue was homogenized, cells were pelleted. Cell lysis was performed (Cell lysis buffer, Cell Signaling Technology, Danvers, MA, USA) and protein electrophoresis initiated. Proteins were transferred to polyvinylidene difluoride (PVDF) membranes, blocked with 5% milk in TBS-Tween, and probed overnight at 4°C with the following primary antibodies: SMAD7 (rabbit anti-mouse, Invitrogen, USA), PTEN (rabbit anti-mouse, Abcam), Foxo3a (rabbit anti-mouse, Cell signaling Technology), phospho-AKT (ser) (rabbit anti-mouse, Cell signaling Technology), AKT (rabbit anti-mouse, Cell signaling Technology), phospho-ERK (44/42) (rabbit anti-mouse, Cell signaling Technology), ERK (rabbit anti-mouse, Cell signaling Technology). Antibody binding was visualized by chemiluminescence (Super-Signal West Pico Chemiluminescent, Thermo Scientific, Rockford, IL, USA). Rabbit anti-mouse glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Sigma-Aldrich) was used as an internal loading control and for normalization of protein quantification. Immunoblots were scanned and quantified using ImageJ densitometry software. Recombinant OPN was purchased at RnD systems (USA). The phosphoinositide-3-kinase (PI3-kinase) inhibitor wortmannin was obtained from Sigma-Aldrich.
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!