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PROTEOSTAT^® Protein refolding and aggregation sensing kit

ENZ-51040

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Identify Optimal Protein Refolding Conditions and Maximize Recovery of Functional Protein

Don’t just refold – with integrated workflow, identify conditions that can cause off-pathway protein aggregation with PROTEOSTAT^® dye
Comprehensive set of optimized screening reagents and conditions that facilitate protein refolding.
Rapid microplate screening using Design of Experiments (DOE)-valued approach to identify parameters that are critical to protein formulation.

PROTEOSTAT^® Protein refolding and aggregation sensing kit provides a concerted assay format employing a fractional factorial matrix design that facilitates screening of refolding parameters for a specific protein in different buffers, and a simple, homogenous assay format for monitoring protein aggregation using a proprietary red-emitting aggregation-sensitive molecular rotor dye. Selected samples with low fluorescence readout, thus low aggregation content, can then validated using traditional enzyme activity-based determination of refolding success.

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PROTEOSTAT® Protein refolding and aggregation sensing kit image — Figure 2. Refolding buffer 13B was determined to have a high activity and low fluorescence, as shown in Figure 1, and was selected to evaluate excipient effects on lysozyme refolding. Excipients were individually added into the refolding buffer with GSH/GSSG environment, and analyzed for aggregation and activity, as in Figure 1. The results indicated that MgCl₂ was the best at promoting refolding in refolding buffer 13B, with minimal tendency to aggregate.

Product Details

Application Notes	PROTEOSTAT^® Protein refolding and aggregation sensing kit provides a fractional factorial matrix design to facilitate screening of refolding parameters for a specific protein in different buffers, and a simple, homogenous assay format for monitoring protein aggregation using a red-emitting aggregation-sensitive dye.
Contents	PROTEOSTAT^® Detection Reagent, 22 µl 10X PROTEOSTAT^®Assay Buffer, 1 x 2 ml 2X Refolding Buffers 1-15, 10 ml each Lysozyme standard, 10 mg Denaturant: 6M Guanidine Hydrochloride (GdnHCl), Tris-HCl (pH 8), 14 ml 100 mM DTT, 1 ml GSH, 46.1 mg GSSG, 18.4 mg 10 mM PEG, 1 ml 500 mM EDTA, 1 ml 400 mM CaCl₂, 1 ml 400 mM MgCl₂, 1 ml 1 M NaCl, 1 ml 0.1% Tween-20, 1 ml 1.2 M Sucrose, 1 ml α-Cyclodextrin (200 mg/ml), 1 ml
Quality Control	Using the procedure described in the manual, denatured lysozyme is refolded to compare the relative fluorescence in different buffers.
Quantity	For 192 reactions
Technical Info / Product Notes	The PROTEOSTAT^®Protein refolding and aggregation sensing kit is a member of the PROTEOSTAT^® product line, reagents and assay kits that have been extensively tested in protein aggregation applications. PROTEOSTAT^® reagents and kits are optimal for demanding protein analyses involving aggregation using microscopy, flow cytometry, microplate readers and HCS/HTS, where consistency and reproducibility are required.

Handling & Storage

Use/Stability	With proper storage, the kit components are stable for one year from date of receipt.
Handling	Protect from light. Avoid freeze/thaw cycles.
Long Term Storage	-20°C
Shipping	Dry Ice