Adipocyte SIK2/SIK3 Activity Assay

Lysates (adipocytes 10–20 μg protein, adipose tissue 100–200 μg protein) were incubated with SIK2 or SIK3 antibodies coupled to protein G-Sepharose (GE Healthcare, Little Chalfont, UK), and phosphotransferase activity towards the peptide substrates HDAC5tide or Sakamototide (200 μmol/l, GL Biochem, Shanghai, China) was measured as described previously [25 (link), 32 (link)]. One unit of activity (U) was defined as that which catalysed the transfer of 1 nmol ³²P/min to the substrate.

Free full text: Click here

Säll J., Pettersson A.M., Björk C., Henriksson E., Wasserstrom S., Linder W., Zhou Y., Hansson O., Andersson D.P., Ekelund M., Degerman E., Stenkula K.G., Laurencikiene J, & Göransson O. (2016). Salt-inducible kinase 2 and -3 are downregulated in adipose tissue from obese or insulin-resistant individuals: implications for insulin signalling and glucose uptake in human adipocytes. Diabetologia, 60(2), 314-323.

Publication 2016

protein G-Sepharose Sakamototide

Adipocytes Adipose tissue Antibodies Peptide Phosphotransferase Protein Protein g Sepharose

Corresponding Organization :

Other organizations : Lund University, Karolinska Institutet

Top 5 similar protocols

Variable analysis

independent variables

SIK2 antibodies
SIK3 antibodies

dependent variables

Phosphotransferase activity towards the peptide substrates HDAC5tide or Sakamototide

control variables

Protein G-Sepharose (GE Healthcare, Little Chalfont, UK)
HDAC5tide or Sakamototide (200 μmol/l, GL Biochem, Shanghai, China)
Lysates (adipocytes 10–20 μg protein, adipose tissue 100–200 μg protein)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!