Universal Linkage System (ULS™): A versatile, chemical labeling technology.

The proprietary ULS™ technology provides the basis of Kreatech's broad range of labeling-based applications for FISH, arrayCGH, miRNA microarrays, gene expression microarrays, protein arrays and others. ULS labeling is based on the stable binding properties of platinum(II) to nucleic acids and proteins [1-3]. The ULS molecule consists of a monofunctional platinum complex coupled to a detectable molecule of choice. The platinum atom forms a coordinative bond, firmly coupling the detectable molecule to the biomolecule of choice: DNA, RNA or protein. ULS labels DNA and RNA by binding to N-7 of guanine. In proteins, ULS binds to sulfur-containing side chains of methionine, and cysteine, and to a nitrogen atom in histidine (see figures below).

Watch a short animation of the ULS.

Schematic Overview of the ULS Technology

Universal Linkage System (ULS™): A versatile, chemical labeling technology.

The proprietary ULS™ technology provides the basis of Kreatech's broad range of labeling-based applications for FISH, arrayCGH, miRNA microarrays, gene expression microarrays, protein arrays and others. ULS labeling is based on the stable binding properties of platinum(II) to nucleic acids and proteins [1-3]. The ULS molecule consists of a monofunctional platinum complex coupled to a detectable molecule of choice. The platinum atom forms a coordinative bond, firmly coupling the detectable molecule to the biomolecule of choice: DNA, RNA or protein. ULS labels DNA and RNA by binding to N-7 of guanine. In proteins, ULS binds to sulfur-containing side chains of methionine, and cysteine, and to a nitrogen atom in histidine (see figures below).

Watch a short animation of the ULS.

Schematic Overview of the ULS Technology

General:

• Chemical labeling method (no enzymes required)
• Easy to control reaction Conditions resulting in optimal control over the degree of labeling which is reproducible
• Easily scalable to the amount of sample to be labeled
• Stable in aqueous solutions (enabling automation) and stored at 4°C

Nucleic Acid Labeling

• Uniform labeling of native DNA, RNA and oligonucleotides as well as enzymatically amplified nucleic acids (e.g. PCR, WGA, aRNA products)
• Size independent labeling of nucleic acids; e.g. ranging from miRNA (20mer) to genomic DNA (>20kb)
• Suitable for the labeling of 3' / 5' modified nucleic acids (e.g. plant and germ-line miRNA)
• Suitable for the labeling of native bacterial mRNA
• Efficiently labels nucleic acids from archival material such as DNA/RNA isolated from formalin-fixed paraffin-embedded (FFPE) tissue
• Optimal removal of unreacted label using KREApure^TM columns

Protein Labeling

• Labels methionine, cysteine and histidine containing proteins and peptides
• High coverage of the proteome (>98%) required for labeling complex protein mixtures
• Labels proteins from any type of sample, like cell lysate, plasma, serum, and even proteins extracted from formalin-fixed paraffin-embedded (FFPE) tissue
• Low chance of interference with epitope recognition or protein interaction domains
• Optimal removal of unreacted label using ULS-Trap^TM columns
• Labels independent of pH and is compatible with many salts and buffers including Tris and glycine

General:

• Chemical labeling method (no enzymes required)
• Easy to control reaction Conditions resulting in optimal control over the degree of labeling which is reproducible
• Easily scalable to the amount of sample to be labeled
• Stable in aqueous solutions (enabling automation) and stored at 4°C

Nucleic Acid Labeling

• Uniform labeling of native DNA, RNA and oligonucleotides as well as enzymatically amplified nucleic acids (e.g. PCR, WGA, aRNA products)
• Size independent labeling of nucleic acids; e.g. ranging from miRNA (20mer) to genomic DNA (>20kb)
• Suitable for the labeling of 3' / 5' modified nucleic acids (e.g. plant and germ-line miRNA)
• Suitable for the labeling of native bacterial mRNA
• Efficiently labels nucleic acids from archival material such as DNA/RNA isolated from formalin-fixed paraffin-embedded (FFPE) tissue
• Optimal removal of unreacted label using KREApure^TM columns

Protein Labeling

• Labels methionine, cysteine and histidine containing proteins and peptides
• High coverage of the proteome (>98%) required for labeling complex protein mixtures
• Labels proteins from any type of sample, like cell lysate, plasma, serum, and even proteins extracted from formalin-fixed paraffin-embedded (FFPE) tissue
• Low chance of interference with epitope recognition or protein interaction domains
• Optimal removal of unreacted label using ULS-Trap^TM columns
• Labels independent of pH and is compatible with many salts and buffers including Tris and glycine

Small molecules are labeled as efficiently as large molecules.

 

ULS labeling is independent of nucleic acid fragment length. (A) Molecules of different fragment lengths were labeled, and the Degree of Labeling was determined. (B) Total RNA was labeled, and separated in 2 fractions, i.e. small RNA, and total RNA without small RNA. Shown are the Degree of Labeling values of the individual fractions.

Labeling of nucleic acids using ULS allows precise control over the degree of labeling.

Small molecules are labeled as efficiently as large molecules.

 

ULS labeling is independent of nucleic acid fragment length. (A) Molecules of different fragment lengths were labeled, and the Degree of Labeling was determined. (B) Total RNA was labeled, and separated in 2 fractions, i.e. small RNA, and total RNA without small RNA. Shown are the Degree of Labeling values of the individual fractions.

Labeling of nucleic acids using ULS allows precise control over the degree of labeling.

Formalin-fixed paraffin-embedded extracted protein.

Protein extracted from FFPE material can be labeled using ULS directly, but not using conventional amine-directed labeling technologies. Shown on the left are the in-solution staining using Dy647-ULS and Dy647-NHS, respectively, and the in-gel staining of the protein content using Coomassie Brilliant Blue (CBB) of the Dy647-labeled samples. 

ULS labeling is compatible with generally used reagents:

• buffers: Tris/glycine, PBS, carbonate etc.
• detergents: NP40, TritonX-100, Tween-20/80 etc.
• cations/anions: Mg2+, Na+, K+, Ca2+, EDTA, EGTA, Cl-, SO42-, PO43- etc

Protein domain interactions.

Since ULS target amino acids differ from NHS labeling, ULS labeling rather allows for labeling outside the protein-protein interaction domains, thereby largely reducing the risk of blocking protein-protein interaction.

Formalin-fixed paraffin-embedded extracted protein.

Protein extracted from FFPE material can be labeled using ULS directly, but not using conventional amine-directed labeling technologies. Shown on the left are the in-solution staining using Dy647-ULS and Dy647-NHS, respectively, and the in-gel staining of the protein content using Coomassie Brilliant Blue (CBB) of the Dy647-labeled samples. 

ULS labeling is compatible with generally used reagents:

• buffers: Tris/glycine, PBS, carbonate etc.
• detergents: NP40, TritonX-100, Tween-20/80 etc.
• cations/anions: Mg2+, Na+, K+, Ca2+, EDTA, EGTA, Cl-, SO42-, PO43- etc

Protein domain interactions.

Since ULS target amino acids differ from NHS labeling, ULS labeling rather allows for labeling outside the protein-protein interaction domains, thereby largely reducing the risk of blocking protein-protein interaction.