As described previously,16 (link) liver tissues from mice fasted for 16 h were extracted, quantified via BCA assay, and equivalent quantities of protein were separated via SDSPAGE (Thermo Scientific, Waltham, MA) and transferred to nitrocellulose membranes. The primary antibodies used were p-AKT, AKT, p-Foxo1 S256, Foxo1, and actin from Cell Signaling Technology (Danvers, Massachusetts) and PI3K-p85, AR (N20) from Santa Cruz Biotechnology (Dallas, Texas), Glucose-6-phosphatase catalytic subunit (G6PC) from Novus Biotechnologicals (Centennial, CO), and phosphoenolpyruvate carboxykinase (PEPCK) from Abcam (Cambridge, MA) all 1:1000 dilutions. The blots were then placed in secondary antibodies (goat anti mouse or goat anti-rabbit, BioRad), and detected using enhanced chemiluminescence (Perkin Elmer Life Sciences, Boston, MA) or Odyssey CLx Imaging System (LI-COR Biosciences, Lincoln, NE).
Phosphoenolpyruvate carboxykinase pepck
Manufactured by Abcam
Sourced in United States
Phosphoenolpyruvate carboxykinase (PEPCK) is an enzyme involved in gluconeogenesis, the metabolic pathway that generates glucose from non-carbohydrate precursors. PEPCK catalyzes the conversion of oxaloacetate to phosphoenolpyruvate, which is a key step in this process.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey clx imaging system, by LI COR (1 mentions)
Pfoxo1 s256, by Cell Signaling Technology (1 mentions)
Sds page, by Thermo Fisher Scientific (1 mentions)
Foxo1, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Phospho akt2 ser474, by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Anti total akt antibody, by Cell Signaling Technology (1 mentions)
Glyceraldehyde 3 phosphate dehydrogenase, by Santa Cruz Biotechnology (1 mentions)
C ebp homologous protein chop, by Cell Signaling Technology (1 mentions)
Phospho eif2a, by Cell Signaling Technology (1 mentions)
Total jnk, by Cell Signaling Technology (1 mentions)
Uncoupling protein 1 ucp1, by Santa Cruz Biotechnology (1 mentions)
Total eif2a, by Cell Signaling Technology (1 mentions)
Hrp conjugated secondary antibody, by Bio-Rad (1 mentions)
2 protocols using phosphoenolpyruvate carboxykinase pepck
1
Western Blot Analysis of Hepatic Proteins
2
Western Blot Analysis of Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoblots were done as previously described (Jornayvaz et al. 2012) . Membranes were incubated overnight with primary antibodies for phospho-Akt2 (Ser 474 ) (Cell Signaling Technology, Danvers, MA, USA), phosphoenolpyruvate carboxykinase (PEPCK; Abcam, Cambride, MA, USA), pyruvate carboxylase (PC; Abcam), uncoupling protein 1 (UCP1; Santa Cruz Biotechnology), C/EBP homologous protein (CHOP; Cell Signaling Technology), IgH chain binding protein (BIP; Cell Signaling Technology), phospho-eIF2a (Cell Signaling Technology), or phospho-JNK (Cell Signaling Technology). After further washings, membranes were incubated with HRP-conjugated secondary antibody (Bio-Rad) and visualized by ECL substrate (Pierce, Rockford, IL, USA). Membranes were stripped and reblotted with anti-total Akt antibody (Cell Signaling Technology), total eIF2a (Cell Signaling Technology), total JNK (Cell Signaling Technology), or glyceraldehyde-3-phosphate dehydrogenase (Santa Cruz Biotechnology). Bands were then quantified using ImageJ (National Institutes of Health, Bethesda, MD, 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!