AMPK Methods and Protocols

Aspirate the supernatant, and the sediment is the lysosome fraction. The purified lysosome can be used for IB by mixing with te ...

Immunofluorescent Analysis of Lysosomal Localization of AXIN For monitoring dynamic translocation of AXIN onto the sur- face of ...

Sonicate the lysates (see Note 34), and centrifuge at 20,000gat 4C for 15 min. Incubate the centrifuged supernatants with ant ...

Each reconstitution reaction requires light organelles from two 15-cm dishes of MEFs. Resuspend the light organelles with 500μL ...

Light organelles of two 15-cm dishes of MEFs for each reac- tion are resuspended with 50μL of fractionation buffer con- taining ...

Open the abdomen to expose the organs in the peritoneal cavity and tightly clamp the edge of liver with the prefreezed clamp fo ...

We strongly recommend that the mouse be housed individually particularly during sacrifice. The lysis buffer can be prepared in ...

the complete DMEM medium directly or administered into animals at chosen doses. It should be noted that AMPK could be artificia ...

8% (v/v) formaldehyde directly into the well containing a similar volume of DMEM medium. The parameters, including PMT voltage, ...

References Hardie DG, Ross FA, Hawley SA (2012) AMPK: a nutrient and energy sensor that main- tains energy homeostasis. Nat Rev ...

Chapel A, Kieffer-Jaquinod S, Sagne C, Verdon Q, Ivaldi C, Mellal M, Thirion J, Jadot M, Bruley C, Garin J, Gasnier B, Journet ...

Chapter 26 Manipulation and Measurement of AMPK Activity in Pancreatic Islets Aida Martinez-Sanchez, Marie-Sophie Nguyen-Tu, Isa ...

such as glucose-stimulated insulin secretion, growth, or apoptosis, remains controversial [5, 6]. Moreover, recent studies sugge ...

1.1 Isolation and Maintenance of Pancreatic Islets We provide a brief description of the method based in collagenase injection i ...

2 Materials 2.1 Isolation and Maintenance of Pancreatic Islets Dissection tools: Surgical scissors (blunt/blunt and sharp/ blun ...

Nontreated 24-well plates. Bicinchoninic acid protein assay reagent from Pierce. Synthetic SAMS peptide (HMRSAMSGLHLVKRR). [γ-^ ...

ECL Prime Western. Re-blot plus strong antibody stripping solution (Millipore). 2.4 Preparation of Islet RNA for RNA- seq Iso ...

Probe sonicator (Ultrasonic homogenizer). Insulin Ultra-sensitive kit (62IN2PEH, Cisbio). 384 wells, low volume plates (white). ...

Incubate the pancreas in a water bath at 37C for exactly 10 min (seeNote 6). Add 15 ml of RPMI and shake vigorously for ~20 s. ...

Handpick islets (seeNote 9) into a 1.5 ml tube, spun for 2 min at 200g, and resuspend in a small volume (50–100μl per conditio ...