Antibody Mega Labeling, Conjugation Kit, MaxLight(TM) Long-Arm Biotin

Product Overview

• Article Name: Antibody Mega Labeling, Conjugation Kit, MaxLight(TM) Long-Arm Biotin
• Biozol Catalog Number: USB-383575
• Supplier Catalog Number: 383575
• Alternative Catalog Number: USB-383575-3X0.1
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

This kit is supplied with Long-Arm Biotin that has been pre-activated for direct use, as well as reagents sufficient for approximately 3 labeling reactions of 0.1-2mg each of antibody or other protein. Each kit includes 6 filtration tubes for desalting and thus eliminates the need for dialysis. The whole procedure is simple and can be completed in 90 minutes. Principle: The NH2-Reactive Long-Arm Biotin reacts with primary amines (N-terminal and the side chain of lysine residues) in antibodies and other proteins, forming a stable amide bond between the antibody/protein and biotin. The long-arm (spacer arm) reduces steric hindrance when several biotinylated molecules bind with avidin/streptavidin, effectively optimizing labeling and detection results. Kit Components: 383575A: Activated Long-Arm Biotin, 3 x 0.17mg 383575B: Labeling Buffer, 1 x 10ml 383575C: DMF (Dimethylformamide), 1 x 500ul 383575D: Filtration tube (6 tubes with 0.5ml capacity per tube) Materials Required But Not Supplied: Adjustable high-precision transfer pipets (10-1000 uL) Incubator (37C) Centrifuge (capable of speed up to 12,000 x g). Storage and Stability: Store kit at 4C. Unopened kit can be stored at 4C for one year before opening and should be used before this expiration date. For maximum recovery of product, centrifuge the original vials prior to removing the cap. Properties of Long-Arm Biotin Molecular Weight: 568 g/mol Molecular Formula: C26H41N5O7S Spacer Arm Length: 30.5 A Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Coupling Procedure: The following procedure is based on labeling of 1mg of antibody/protein. Adjust volume of Activated Biotin Solution relative to the actual amount of antibody/protein to be labeled. 1. Mix 1mg of protein sample and corresponding volume of labeling buffer to filtration tube to make the total volume equal to 0.5ml. Centrifuge at 12,000 x g for 10 minutes. Note: The maximum volume capacity of the filtration tube is 0.5ml. If the protein concentration in the sample is low, the sample can be concentrated first using centrifugal ultrafiltration (use filters with appropriate molecular weight cut-off). 2. Add 13.3ul of Activated Long-Arm Biotin Solution and appropriate volume of Labeling Buffer to the filtration tube to make the final concentration of the protein solution equal to 2mg/ml. Mix thoroughly using a pipet and incubate the tube for 30 minutes at 37C. 3. Centrifuge at 12,000 x g for 10 minutes. 4. Add appropriate volume of Labeling Buffer to the filtration tube to make the total volume equal to 0.5ml. Mix thoroughly using a pipet and centrifuge at 12,000 x g for 10 minutes. 5. Repeat step 4. 6. Add 0.2ml of Labeling Buffer to the filtration tube and mix thoroughly using a pipet. Invert the filtration tube into another centrifugal tube. Centrifuge at 6,000 x g for 10 minutes. 7. Collect the solution in the centrifugal tube, this liquid will contain the biotinylated antibody/protein.