MONMOUTH JUNCTION, NJ – Kinase research on identifying druggable kinome modulating activity using small molecule kinase inhibitors, cyclic inhibitors and binding non-ATP sites requires proper sample preparation. Enrichment of kinases using KinaSorb™ provides samples ready for biochemical and cell-based assays.

KinaSorb™ is a new reagent kit used for the enrichment and isolation of kinases. In a patent pending process, ATP – the common substrate for kinases is reversibly immobilized to metallic oxide particles, producing a single-use highly efficient enrichment method for kinase & other ATP binding proteins. The standard prep protocol starts with 100 µl of clarified cellular extracts, or approximately 2 mg total cellular extract protein, but the process can be scaled up or down to accommodate different sample volumes and protein concentrations. The methods are reasonably specific to ATP; little phosphodiesterase activity (an enzyme which binds to a structurally similar adenosine containing substrate – CAMP), was measured in the eluant after KinaSorb™ treatment. The kit includes all necessary reagents for immediate use.

Kinase assay detection kits benefit from enrichment protocols from samples of purified enzymes, cell lysates, tissue homogenates, cell extracts. The enriched kinases are subsequently used for various applications including:

• Kinase assay protocol development of PKA, PKC and PTK from heart, spleen, lymphocytes, skin, lung & fibroblasts (various 3T3 lines), promyelocytic cells (HL-60 and U937) and neuron-like cell lines (PC12).
• Kinase activity measurement and probe labeling
• Method development of buffer composition, ATP binding proteins, quantification of enriched kinases eg: multiple-reaction monitoring mass spectrometry (MRM-MS) assays
• Kinase experiments on cell lines eg: A375, A549, HUH7, MDA-MB-231, Jurkat, Ramos, THP1, Hel, HeLa, HL60, NIH3T3, OVCAR3. Kinase experiments on tissues: uman PBMC, mouse brain, mouse liver, mouse spleen, mouse lung, rat brain, rat white blood cells.

Characteristics of KinaSorb™

• Ratio of Total Kinase Activity* relative to total protein content increases 300% or more
• Simple bind, wash and elute protocol
• Mild elution maintains tertiary structure and simple transfer to secondary analysis
• ATP binding protein recoverable, ~200 ug
• Up to 4X concentration of kinase volume, relative to starting volume
• 60 minute, scaleable protocol compatible with functional assays, electrophoresis and mass spectrometry
• Phosphatase (neutral & alkaline) activity not detectable
• ?Improves protein normalization when comparing heterogeneous tissues

For more information visit: KinaSorb™*
http://www.biotechsupportgroup.com/node/72

About Biotech Support Group LLC

Biotech Support Group LLC is a leading provider of genomics and proteomics sample preparation products and enrichment reagent kits as well as integrated biotechnology services for life sciences research, biomarker and drug discovery. Based in New Jersey, it’s principal products include: AlbuSorb™& AlbuVoid™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, HemogloBind™ & HemoVoid™ for hemoglobin removal, NuGel™ for functional & chemical proteomics, and ProCipitate™ & ProPrep™ for nucleic acid isolation. For
more information, go to www.biotechsupportgroup.com

CONTACT:
Dr. Swapan Roy & Matthew Kuruc
Biotech Support Group LLC
1 Deer Park Drive, Suite M
Monmouth Junction NJ 08852
732-274-2866 Worldwide
800-935-0628 North America

sales@biotechsupportgroup.com
http://www.biotechsupportgroup.com

*For research use only