Ubiquitin Linkage Screening Kit II is designed for two purposes: 1) to identify the ubiquitin lysine residue that is used to link the polyubiquitin chain conjugated on a protein substrate; 2) to determine whether a protein substrate is polyubiquitinated or conjugated with multiple monoubiquitin.

* The supplied UBE1 is sufficient for setting up 50 x 20 µl reactions. Each Ub mutant is sufficient for setting up 10 X 20 µl reactions.
Description

Ubiquitin Linkage Screening Kit II is designed for two purposes: 1) to identify the ubiquitin lysine residue that is used to link the polyubiquitin chain conjugated on a protein substrate; 2) to determine whether a protein substrate is polyubiquitinated or conjugated with multiple monoubiquitin.

A typical 20 µl reaction contains 100 nM UBE1, 2 µM E2 (not supplied), 50 µM Ub or Ub mutants, 4 mM ATP, 2 µM E3 (not supplied) and 2 µM substrate (no supplied) in the ubiquitination buffer.

Components

• 20X UBE1 (2 µM)..................................50 µl
• 10X 6xHis-Ub (500 µM)........................20 µl
• 10X 6xHis-Ub(K0) (500 µM)................20 µl
• 10X 6xHis-Ub(K6R) (500 µM).............20 µl
• 10X 6xHis-Ub(K11R) (500 µM)...........20 µl
• 10X 6xHis-Ub(K27R) (500 µM)...........20 µl
• 10X 6xHis-Ub(K29R) (500 µM)...........20 µl
• 10X 6xHis-Ub(K33R) (500 µM)...........20 µl
• 10X 6xHis-Ub(K48R) (500 µM)...........20 µl
• 10X 6xHis-Ub(K63R) (500 µM)...........20 µl
• 10X ATP (40 mM)................................125 µl
• 10X Ubiquitination Buffer.........................1 ml