Buccutite™ FOL-Dye 650
Our Buccutite™crosslinking technology provides the most convenient and effective crosslinking method to link two biomolecules with a high conjugation yield. Our method uses one pair of crosslinkers: Buccutite™ MTA and Buccutite™ FOL. MTA is added to one molecule, while FOL is added to another molecule. The cross-linking reaction is initiated by mixing Molecule-1-Buccutite ™ MTA and Molecule-2-Buccutite ™ FOL. This crosslinking reaction occurs under extremely mild and neutral conditions without any catalyst required. It is robust and efficient. Many of our customer have requested us to offer the stand-alone Buccutite™ MTA and Buccutite™ FOL reagents to expand the application of Buccutite™crosslinking technology. This Buccutite™ FOL reagent is used to determine the number of FOL groups of the Molecule-2-Buccutite ™ FOL. The number of FOL linkers provides an important parameter to to optimize crosslinking process.
![Buccutite™crosslinking technology provides the most convenient and effective crosslinking method to link two biomolecules with a high conjugation yield. Our method uses one pair of crosslinkers: Buccutite™ MTA and Buccutite™ FOL. MTA is added to one molecule, while FOL is added to another molecule. The cross-linking reaction is initiated by mixing Molecule-1-Buccutite ™ MTA and Molecule-2-Buccutite ™ FOL. This crosslinking reaction occurs under extremely mild and neutral conditions without any catalyst required. It is robust and efficient.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fbuccutite-fol-dye-650%2Ffigure-for-buccutite-fol-dye-650_YYaLe.jpg&w=640&q=75)
![Buccutite™crosslinking technology provides the most convenient and effective crosslinking method to link two biomolecules with a high conjugation yield. Our method uses one pair of crosslinkers: Buccutite™ MTA and Buccutite™ FOL. MTA is added to one molecule, while FOL is added to another molecule. The cross-linking reaction is initiated by mixing Molecule-1-Buccutite ™ MTA and Molecule-2-Buccutite ™ FOL. This crosslinking reaction occurs under extremely mild and neutral conditions without any catalyst required. It is robust and efficient.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fbuccutite-fol-dye-650%2Ffigure-for-buccutite-fol-dye-650_YYaLe.jpg&w=640&q=75)
![Buccutite™crosslinking technology provides the most convenient and effective crosslinking method to link two biomolecules with a high conjugation yield. Our method uses one pair of crosslinkers: Buccutite™ MTA and Buccutite™ FOL. MTA is added to one molecule, while FOL is added to another molecule. The cross-linking reaction is initiated by mixing Molecule-1-Buccutite ™ MTA and Molecule-2-Buccutite ™ FOL. This crosslinking reaction occurs under extremely mild and neutral conditions without any catalyst required. It is robust and efficient.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fbuccutite-fol-dye-650%2Ffigure-for-buccutite-fol-dye-650_YYaLe.jpg&w=128&q=25)
Example protocol
PREPARATION OF STOCK SOLUTION
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Buccutite™ FOL-Dye 650 stock solution (10 mM):
Add 200 uL DMSO to Buccutite™ FOL-Dye 650 vial to prepare 10 mM stock solution. Note: The Buccutite™ FOL-Dye 650 stock solution should be stored at -20 °C after preparation and stable for 2 months if avoid repeated freeze-thaw cycles.
SAMPLE EXPERIMENTAL PROTOCOL
FOL Sample Preparation
- Use 100 ug FOL-modified sample (for example: antibody or other protein modified with MTA group, the MW should be above 15,000).
- Adjust the volume to 100 uL with PBS.
Run FOL Assay
- Add 10 uL 10 mM Buccutite™ FOL-Dye 650 stock solution to FOL sample solution.
- Keep the reaction mixture at room temperature and rotate or shake it for 60 minutes.
- Prepare spin column (Cat#60500) for sample purification.
- Load the reaction mixture to a spin column with a clean collecting tube. After all the solution loaded to the column, add 10 uL PBS to the top and centrifuge the column for 5 minutes at 1,000 x g.
- Collect the solution with a collecting tube.
- Measure the absorption spectra with 0.5 mL Quartz Cuvette or Nanodrop. Note: Dilute the elution by 5 - 10 folds with PBS, measure the absorption spectrum from 800 nm to 250 nm, or only read the absorbance number at 280 nm and 654 nm.
- Calculate FOL # (moles of FOL / mole of molecule) with the following equation.
FOL # = (A654 / 250000) / {(A280 - 0.09 X A654) / EC}
A280: absorbance of the elution at 280 nm
A654: absorbance of the elution at 654 nm
EC: Extinction Coefficient of the sample (M-1cm-1)
Calculators
Common stock solution preparation
Table 1. Volume of Water needed to reconstitute specific mass of Buccutite™ FOL-Dye 650 to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 87.989 µL | 439.947 µL | 879.894 µL | 4.399 mL | 8.799 mL |
5 mM | 17.598 µL | 87.989 µL | 175.979 µL | 879.894 µL | 1.76 mL |
10 mM | 8.799 µL | 43.995 µL | 87.989 µL | 439.947 µL | 879.894 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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