Thiol kits

If your biomolecule does not have a free thiol group, it will require activation to generate a free thiol group prior to conjugation with the maleimide label:

1. Protein Thiolation kit

Designed to facilitate the conjugation of proteins to our maleimide-activated labels. Simply add your protein solution to the lyophilized thiolation reagents to activate your protein. These kits are designed to activate 2mg or 5mg of your protein. Each thiolation kit is also supplied with a thiolation detection kit provided as a positive test for protein thiolation, giving you the option to confirm the chemistry is working.

2. Thiol quantification kit

The thiol quantification assay kit allows you to quantify the amount of thiol/sulfhydryl (-SH) groups in samples of thiolated proteins or antibodies. Quantifying the number of free thiols in a sample can be useful in a number of applications and allows for a more controlled conjugation reaction. The assay is in a convenient 96-well plate format and contains all the necessary reagents required.

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  1. When and why would you quantify the number of free thiols in a sample?
    Quantifying the number of free thiols in a sample can be useful in a number of applications. For example, if the final purpose is the conjugation of a thiolated molecule to a maleimide – activated label, quantifying the number of free thiols on the molecule is highly recommended in order to have a more controlled conjugation reaction.
  2. What is the minimum thiol concentration detected with the assay?
    A concentration of 1.5 µM of thiols can be easily detected.
  3. How do I remove the thiolating agent before testing?
    Dialysis of thiolated antibodies/ proteins is not recommended due to the consequent big dilution. The thiolating reagent can be  removed using a small size-exclusion/gel filtration gravity or spin column. Consult manufacturers’ instructions for use and select resin for your needs.
  4. My buffer is outside the recommended ranges, what should I do?
    If the buffer is too concentrated, dilute the sample to within buffer range. If the sample cannot be diluted further, the buffer could alternatively be exchanged using a size exclusion/gel filtration gravity or spin column.
  5. What if the measured absorbance of the sample is out of the range set by the calibration curve?
    Concentrate or dilute the sample in order to be within the linear range of the calibration curve.

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