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    OptiResc Protein Aggregate Solubilization Kit

    $279.00$789.00

    This OptiResc™ Protein Aggregate Solubilization Kit contains a systematically varied array of buffers (from pH 3 to pH 10) and a series of solubility enhancers (salts, amino acids, sugars, polyols, reducing reagents) that help to determination conditions under which a particular aggregated protein sample dissolves.

    Additional Discount: Order 12 or more Kits, an additional 5% discount will be applied at checkout.


    Discount Per Quantity

    Quantity 12 +
    Price $265.05$749.55
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    Catalog: OptiResc

    Why OptiResc ?

    Proteins aggregate for a variety of reasons. Fortunately, reversibly aggregated protein samples can be brought back into solution by adjusting pH, ionic strength and additive content. This OptiResc Protein Aggregate Solubilization Kit contains a systematically varied array of buffers (from pH 3 to pH 10) and a series of solubility enhancers (salts, amino acids, sugars, polyols, reducing reagents) that help to determination conditions under which a particular aggregated protein sample dissolves.

    • Quickly evaluate solubilization behavior as a function of pH, salt and additives
    • Assay more than 90 different conditions in parallel and label-free
    • Simply mix and observe with a microscope, no further handling required

    One OptiResc kit contains all consumable materials to identify solution conditions of up to six (6) different, reversibly aggregated protein samples.


    View the reagent listing and protocol in OptiResc™ Quick Start Guide

    Applications

    The OptiResc Solubization kit is designed to bring “crashed out” protein sample back into solution. If the sample has reversibly aggregated and visibly precipitated the systematically varied array of buffers in the kit can help to determine the right buffer to dissolve the aggregated protein sample.

    Proteins aggregate. This undesired process is often encountered when changing buffers, after proteolytic cleavage of an epitope tag or just over a period of time. Reversibly aggregated protein solutions can be ‘brought back’ into solution by dissolving them into the correct buffer (see this video). The OptiResc Protein Aggregate Solubilization Kit contains a systematically varied array of buffers (from pH 3 to pH 10) and a series of solubility enhancers (salts, amino acids, sugars, polyols, reducing reagents) that help to determination conditions under which a particular aggregated protein sample dissolves.

    The benefits:

    • Quickly evaluate solubilization behavior as a function of pH, salt and additives
    • Assay more than 90 different conditions in parallel and label-free
    • Simply mix and observe with a microscope, no further handling required

    Watch this short movie to see how it works:

    Technology

    This is how protein aggregate solubilization with OptiResc works

    The OptiResc Kit contains a systematically varied array of buffers (from pH 3 to pH 10) and a series of solubility enhancers (salts, amino acids, sugars, polyols, reducing reagents) that help to determination conditions under which a particular aggregated protein sample dissolves.

    The visibly aggregated protein sample is mixed with aliquots from each of the OptiResc reagents and the results are inspected under a microscope. Clear drops (dissolved protein) indicate the best solutions to de-aggregate the protein sample.

    optiresc1

    PROTOCOL: IDENTIFY CONDITIONS THAT SOLUBILIZE REVERSIBLY AGGREGATED PROTEIN SAMPLES:

    Material: aggregated protein sample (ca. 50 uL total for each protein sample to be tested) – up to six different protein samples can be applied with one kit (3 Inspection Plates with 2 x 96 OptiResc experimental wells).

    Protocol for two aggregated protein samples:

    1. Distribute 50 uL aliquots from wells from OptiResc Reagent plate wells into large reservoir wells of OptiResc Inspection Plate (all 96 wells).
    2. Transfer 2 x 0.5 uL from each reservoir well of OptiResc Inspection Plate into small wells of OptiResc Inspection plate.
    3. Add 0.5 uL of aggregated protein sample A to each OptiResc experimental well (upper small well A) and add sample B to each OptiResc experimental well (lower well B) within the OptiResc Inspection Plate. Omit sample in position H1 (negative control).
    4. Seal OptiResc Inspection Plate with one Seal Sheet.
    5. Inspect small wells with a microscope, i.e. a dissecting stereo microscope and note the wells that contain precipitate (aggregated protein solution) or clear drops (dissolved protein). If no clear drops are seen, inspect again after 1 hour or 1 day.
    6. Use the OptiResc Reagent Listing to identify solutions that yield clear drops.

    Scale up disaggregation reaction by mixing equal volumes of aggregated protein sample with solution that was identified with the OptiResc kit.

    Data Analysis

    The results of the OptiResc Protein Aggregate Solubilization Screen can be inspected under a stereo microscope: The drops that turn from turbid to clear are the ones that contain a superior buffer since they convert the reversibly aggregated sample into a solubilized sample. This usually takes less than 2 minutes.
    optiresc1

    Scale up

    Prepare larger quantities of the identified formulation and combine with the aggregated protein sample. It is recommended to add small portions of the aggregated protein sample   to a large volume of rescue formulation.

    An example for a small rescue experiment is shown in this short movie:

    OptiSol or OptiResc

    What are the differences between the OptiSol Protein Solubility screening kit and the OptiResc Protein Solubilization kit?

    The starting material for the OptiResc kit is visibly aggregated protein. The protocol asks for dilution of such protein precipitate with the formulation followed by microscopic inspection. Keep in mind that the identification of the formulation that dissolves your precipitated protein is qualitative in nature.

    The starting material for OptiSol kit can be either non-aggregated or aggregated material. Protein aggregates come in different forms, some of them visibly precipitated, others look clear to the eye, even though their microstructure is that of an aggregate, rather than monodisperse particles. The OptiSol kit contains filterplates with molecular weight cutoff filters (30kDa and 200 kDa). This allows you to separate e.g. monomeric from oligomeric protein species and identify the effect of pH, salt, detergent etc. on the aggregation status of the target protein.

    The output of the OptiSol assay is of quantitative nature. The associated Protein Dashboard automatically graphs trendlines through the >90 datapoints, simplifying the analysis of the solubility behavior.

    What tools do I need for the OptiSol vs the OptiResc Kit?

    OptiSol: All you need is a centrifuge that can spin 96-well microtiter plates and an assay to quantify your protein. A protein assay works for purified protein. For protein mixtures a target specific assay makes most sense.

    OptiResc: You really just need a microscope. You’ll need a pipettor to combine the formulation from the reagent plate with the aggregated protein in the inspection plate, observe with the microscope and instantly see which formulation de-aggregates your protein sample.

    Are the solutions the same in both kits, OptiSol and OptiResc?

    Yes, the formulations and the volumes are identical.

    How many samples can I process with one kit? How many “runs” are in each?

    The OptiSol protein solubility screening kit is designed to be run with a single protein sample only. While we have heard from users that have employed the filter plate several times successfully, we don’t recommend it. Multiple use will eventually clog the filter and make the results unreliable.

    The OptiResc kit can be used for 6 different samples. The starting material is visibly aggregated protein solution.

    How labor intensive is the OptiSol kit compared to the OptiResc?

    OptiResc is a lot simpler to set up. No centrifugation. No biochemical assay. Just combine the formulation with the aggregated protein in the inspection plate and observe with a microscope.

    FAQ

    WHAT IS THE COMPOSITION OF THE OPTIRESC SCREENS?

    The OptiResc reagents consist of a combinatorial solution set with a wide variety of pH, buffer types, charged & neutral organic additives and antioxidants. A detailed list of all the components are listed in the OptiResc brochure included in the kit.

    ARE THE OPTIRESC BUFFERS THE SAME AS IN THE OPTISOL KIT?

    Yes, the buffers are the same.

    ARE THE BUFFER CONCENTRATIONS LISTED IN THE DATA SHEET FINAL OR THE STOCKS?

    The concentrations listed are those of the reagents in the wells of the OptiTherm Reagent Plate.

    WHAT IS SOLUBILISIN II?

    Solubilisin II is a proprietary solution that has been shown to promote the solubility and protect proteins from aggregation over a wide range of pH values.

    CAN SOLUBILISIN II USED IN PK STUDIES IN VITRO?

    Solubilisin II has not been tested for in vivo studies and should be used for in vitro studies only. Some of the reagents supplied with the OptiTherm kits may contain low amounts of sodium azide and are hence incompatible with living cells. The OptiTherm kit is for in vitro research use only, not for cell-based or animal research.

    DO YOU HAVE AN MSDS FORM?

    Yes. Contact us at info@solublebioscience.com