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HTRF SEB Supplemented Enzymatic Buffer, 1 Vial

The Supplemented Enzymatic Buffer (SEB) is a proprietary substrate stabilizing component that may be used with KinEASE TK assay to optimize the enzymatic reaction for some tyrosine kinases while maintaining inhibitor properties such as IC50.

For research use only. Not for use in diagnostic procedures.
Part number: 61SEBALB
List price: USD 913.68
Your price:
USD 913.68
USD 913.68 /each

Overview

Some Tyrosine kinase enzymatic activity may require further assay optimization for a sensitive detection of phosphorylation levels. The Supplemented Enzymatic Buffer is a substrate stabilizing buffer that may be used to achieve an optimal enzymatic step prior to detection with the HTRF KinEASE TK kit, while maintaining inhibiting compound pharmacological characteristics.

Specifications

Assay Points
1
Assay Target Type
Buffer
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Shipping Conditions
Shipped Ambient
Therapeutic Area
Metabolism/Diabetes
NASH/Fibrosis
Oncology & Inflammation
Unit Size
1 Vial

Video gallery

Analytical performance

Supplemented Enzymatic Buffer performance

The HTRF KinEASE TK reaction may be run in the presence of SEB, a proprietary substrate stabilizing buffer. This buffer greatly enhances S/B and enables the use of lower kinase concentrations per well. Among validation on other tyrosine kinases, the case below shows the impact of SEB on tests conducted with Abl (Merck Millipore 14-459).

In this SEB titration run, Abl used at 5 ng/well, was incubated 30 min with substrate-biotin (1 µM), and a non-limiting ATP concentration (100 µM) in a final assay volume of 20 µl. SEB concentrations ranging from 0 nM to 50 nM were tested. As shown here, SEB enhances S/B quite significantly.

HTRF SEB Supplemented Enzymatic Buffer 1.svg

 

In this second experiment, Abl was used at concentrations ranging from 0.10 to 10 ng/ml under similar substrate and ATP conditions as previously described, and with either 2.5 or 5 nM SEB. The subsequent S/B improvement at increasing SEB levels makes the use of lower tyrosine kinase concentration possible.

HTRF SEB Supplemented Enzymatic Buffer 2.svg

 

    Tyrosine KinEASE assay
Enzyme Compound With SEB 50 nM Without SEB
Abl Staurosporine
Gleevec
0.108
0.043
-
-
CSK Staurosporine
Nexavar
0.493
4.220
0.417
6.175
AGFR Staurosporine
Iressa
1.069
0.001
0.861
0.002
JAK2 Staurosporine
Sutent
0.00030
0.800
0.00035
0.778
JAK3 Staurosporine
Sutent
0.001
7.639
0.001
4.597
PTK5 Staurosporine
Nexavar
0.039
> 0.1
0.036
> 0 .1

 

Resources

1-1 of 1 Resources
Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

SDS, COAs, Manuals and more Illuminator

SDS, COAs, Manuals and more

Are you looking for technical documents for this product. We have housed them in a dedicated section., click on the links below to explore.

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