One-Step Free Biotin Removal Kit

Introduction

Biotin, also referred to as vitamin H, is a small molecule that is not commonly found in biological samples. It has a remarkable binding affinity for avidin (Ka=10^15) and streptavidin (Ka=10^14), making biotinylated molecules such as proteins or other molecules very useful for non-radioactive detection and purification of target molecules.

Biotinylation is a process used to attach biotin covalently to proteins, nucleic acids, carbohydrates, or other molecules using enzymatic or chemical methods. This creates a biotinylated molecule with biotin attached to it. However, a certain amount of free biotin may remain in the sample after biotinylation, which can cause non-specific binding and result in background noise. Therefore, it is crucial to remove free biotin from the sample for successful downstream applications.

In order to eliminate any excess biotin in a sample, conventional techniques like dialysis or spin columns are often utilized. Unfortunately, these methods have some drawbacks, including loss of the sample due to protein precipitation, multiple buffer exchanges, transfer steps, lengthy protocols, and difficulty with low volume samples or high-throughput automation.

To overcome these challenges, a new and efficient biotin removal system has been developed.

Hygia One-Step Free Biotin Removal Kit uses magnetic beads modified with proprietary chemistry to remove free biotin from sample. The beads can quickly and efficiently remove free biotin from ultra-low volumes of protein/ peptide or DNA/RNA solutions. The beads enable 96 samples to be processed simultaneously in less than 10 minutes.

The beads allow rapid and efficient removal of free biotin from the sample. The procedure is straightforward.

1. Add the beads directly to the sample.

2. Pipette or vortex to capture the free Biotin.

3. Magnetic separation of the beads from the protein, or DNA/RNA solution, while the protein or DNA/RNA remains in the solution.

The easy-to-use magnetic beads significantly improve results over the standard drip column and batch methodologies with minimum protein loss (<10%). Since only a small volume of magnetic beads is used, the final protein concentration of the sample is not significantly decreased.

Features and Advantages

● Simple protocol: No liquid transfer, One-tube, One-step, and one-minute protocol

● Easy-to-use

● Reliable and reproducible results with exceptional >90% recovery for protein (>6 kDa, aprotinin) or DNA/RNA (>25mer dsDNA)

● Effective Cleanup: Remove 95% free Biotin

● Cost-effective: Eliminates columns, filters, and laborious repeat pipetting

● High throughput: Compatible with many different automated liquid handling systems