Tissues were sonicated with RIPA buffer. Protein quantification with BCA assay and 40ug of protein loaded into each well. We used the BioRad XT criterion gel system with MOPS running buffer (#1610788). Proteins were transferred to PVDF membrane and blocked with 5% milk prior to antibody probing. We used the following primary antibodies: pAkt S473 (#8200S), Akt (#8200S), pAMPK T172 (#8208S), AMPK (#8208S), pRPS6 S235 (#8207S), RPS6 (#8207S), pULK S555 (#23988S), and ULK1 (#23988S), all were from Cell Signal and probed overnight at 4 C in TBST. Blots were incubated with secondary HRP antibody for 1 h prior to chemiluminescent treatment and visualization on our BioRad imager. Blots were analyzed with BioRad Image Lab software version 6.1. Raw western blot images are available as Additional file 3 .
Pulk s555
Manufactured by Cell Signaling Technology
Sourced in United States
The PULK S555 is a laboratory instrument designed for the detection and quantification of protein phosphorylation events. It utilizes a sensitive and specific fluorescence-based detection method to measure the level of phosphorylation on specific amino acid residues within target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Pampkt172, by Cell Signaling Technology (2 mentions)
Mops running buffer, by Bio-Rad (1 mentions)
Image lab software version 6, by Bio-Rad (1 mentions)
Pakt s473, by Cell Signaling Technology (1 mentions)
Ns1643, by Alomone (1 mentions)
Procaspase 3, by Cell Signaling Technology (1 mentions)
P21waf, by Cell Signaling Technology (1 mentions)
P mek, by Cell Signaling Technology (1 mentions)
Cyclin e, by Cell Signaling Technology (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
2 protocols using pulk s555
1
Western Blot Profiling of Cell Signaling
2
Investigating Cell Signaling Pathways
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Rabbit Anti AMPK, p-AMPK T172, Pro-Caspase 3, ULK, p-ULK S555, p21WAF, cyclin E, D, & B, MEK, p-MEK, ERK 1/2, p-ERK ½ and pWEE antibodies were purchased from Cell Signaling Technologies, Inc. (Boston, MA, USA). p16INK4A was purchased from Abgent (San Diego, USA, CA). The Kv 11 agonist NS1643 was purchased form Alomone Labs (Jerusalem, Israel).
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