Frequently Asked Questions
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How much of the FabRICATOR do you need to cleave IgG in 50 µl of normal human serum?
If we assume that the antibody concentration in serum is 15 mg/ml you would have to add 10-15 000 units to a sample of 1 ml serum. In 50 µl serum you would have to add 500 – 750 units of FabRICATOR.
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Is it possible to use FabRICATOR in a buffer containing EDTA or HSA?
Yes. FabRICATOR is compatible with EDTA. Make sure your pH is within 6-8.
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Can I cleave Fc-fusion proteins with FabRICATOR?
Yes, FabRICATOR should be able to digest a Fc fusion protein as long as you have not fused the protein too close to the cleavage site. FabRICATOR digests human IgG just below the hinge (…CPAPELLG / GPSVF…), leaving the hinge on the fusion-part.
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Can I use SialEXO for desialylation of both N- and O-glycans?
Yes, SialEXO displays activity on sialic acids with α2-3, α2-6 and α2-8 linked bonds of both N- and O-glycans.
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Does SialEXO require denaturation of the proteins for activity on glycoproteins?
No, SialEXO works both on glycoproteins in native conditions but can also be used on denatured proteins after buffer exchange.
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Does SialEXO contain a purification tag?
Yes, SialEXO contains a 6x His-tag.
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Can I use SialEXO in combination with PNGaseF?
Yes, both enzymes are active under native conditions, preferably use 20 mM Tris pH 7. If denaturing reaction conditions are required for PNGaseF, the reaction with SialEXO needs to be performed first under native conditions, then add PNGaseF and the required buffer.
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Can I use SialEXO in combination with OglyZOR or OpeRATOR?
Yes, SialEXO can be incubated together with OpeRATOR and/or OglyZOR in 20 mM Tris pH 6.8.
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Can OpeRATOR and SialEXO be used in the same reaction?
Yes, the enzymes work in combination and can be used to desialylate and digest an O-glycosylated protein.
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Is removal of sialic acids really needed?
The activity of OpeRATOR is significantly decreased in the presence of sialic acids. We recommend using SialEXO to remove the sialic acids. If this is not an option, then we suggest trying to digest both with and without SialEXO to evaluate the impact for your specific samples and requirements.
Learn more about OpeRATOR™