For extraction of whole cellular proteins, cells were washed twice with ice-cold phosphate buffered saline and then lysed with cell lysis buffer (50 mM Tris-HCl pH 7.4, 1% NP-40, 0.25% Na-deoxycholate, 150 mM NaCl, 1 mM EDTA, 10 mM Na3VO4, 50 mM NaF, 1 mM PMSF, 1 µg/mL aprotinin, 1 µg/mL leupeptin, 1 µg/mL pepstatin) on ice for 30 minutes. Lysates were sonicated, and the cell homogenates were centrifuged at 15,000 g for 10 minutes (4℃). After centrifugation, supernatants were electrophoresed in 10% acrylamide gels and transferred onto nitrocellulose membranes. The proteins were probed overnight with antibodies against JNK, ERK p44/42, p38, NF-κB p65, phospho-JNK, phospho-ERK p44/42, phospho-p38, phospho-NF-κB (Cell Signaling Technology, Danvers, MA, USA) and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The immunoreactive bands were detected with horseradish peroxidase-conjugated secondary antibodies and visualized by enhanced chemiluminescence.
Erk p44 42
Manufactured by Cell Signaling Technology
Sourced in United States
ERK p44/42 is a protein that plays a key role in the mitogen-activated protein kinase (MAPK) signaling pathway. It functions as a regulator of cell growth, division, and differentiation.
Automatically generated - may contain errors
Lab products found in correlation
Nf κb p65, by Cell Signaling Technology (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Phospho nf κb, by Cell Signaling Technology (1 mentions)
Bcl 2, by Abcam (1 mentions)
Ibright fl1000, by Thermo Fisher Scientific (1 mentions)
Phospho ser133creb, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Creb1, by Cell Signaling Technology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
2 protocols using erk p44 42
1
Whole Cellular Protein Extraction and Immunoblotting
2
Quercitrin Modulates Apoptosis Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DPCs (1 × 10 6 cells/dish) were seeded in 100 mm dishes and cultured for 24 h. Quercitrin was treated at concentrations of 1, 10, and 100 nM for appropriate time. The cells were then lysed and total cellular proteins were prepared. 50 μg protein samples were analyzed by Western blotting with corresponding antibodies; Bcl2 (1:1000, Abcam, cambridge, UK), Akt (1:1000, Santa Cruz, CA, USA), Erk (p44/42) (1:1000, Cell Signaling Technology, Danvers, MA, USA), CREB-1 (Cell Signaling Technology, Danvers, MA, USA) GAPDH (1:2000, Santa Cruz, CA, USA), phospho (Ser473)-Akt (1:1000, Cell Signaling Technology, Danvers, MA, USA), phospho (Thr202/Tyr204)-Erk (p44/42) (1:1000, Cell Signaling Technology, Danvers, MA, USA), and phospho (Ser133)-CREB (1:1000, Cell Signaling Technology, Danvers, MA, USA). Western blot was analyzed by chemiluminescence detector iBright FL1000 (Invitrogen, Waltham, MA, USA).
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!