All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: HPRD-10 (for 10ml)

Cat. No.: HPRD-100 (for 100ml)

Cat. No.: HPRD-1k (for 1000ml)

Description

HisPrem™ His-tag Purification Resin (Denaturant-resistant) is a His-tag protein purification medium that can withstand denaturing agents, commonly known as a nickel column. This product is a novel purification medium that can be compatible with strong protein denaturants such as urea and guanidine hydrochloride, allowing for simple, fast, efficient, and highly specific purification of His-tagged proteins under both denaturing and non-denaturing conditions.

Recombinant proteins containing a 6xHis-tag or more consecutive histidine residues are commonly referred to as His-tagged proteins. When a protein sample solution passes through HisPrem™ His-tag Purification Resin (Denaturant-resistant), the histidine residues on the His-tag of the recombinant protein selectively bind to the nickel ions in the HisPrem™ His-tag Purification Resin (Denaturant-resistant), while other proteins do not bind. After washing, the His-tagged recombinant protein can be eluted under non-denaturing or denaturing conditions, resulting in its separation and purification.

This product exhibits strong affinity, high selectivity, and large binding capacity for His-tagged recombinant proteins. It can be used to obtain highly pure His-tagged recombinant proteins from various expression systems, such as bacteria, mammalian cells, insects, and baculoviruses, in a simple, fast, and efficient manner. The purified proteins can be used for structural and functional studies, antibody preparation, protein-protein interactions, protein-nucleic acid interactions, and other research applications.

This product utilizes a highly cross-linked 6% agarose gel as the matrix and employs optimized arm chemistry and nickel ion chelation technology. Compared to similar products on the market, it significantly reduces non-specific protein binding.

This product can tolerate 8 M urea or 6 M guanidine hydrochloride and can be used for the purification of His-tagged proteins under both non-denaturing and denaturing conditions. Reducing agents such as DTT and β-mercaptoethanol can reduce the chelated nickel ions, while chelators such as EDTA/EGTA can cause nickel depletion in this product. Therefore, solutions containing reducing agents and chelators should not be used in protein samples or during the purification process. If protein purification is required under conditions with strong reducing agents like DTT/β-mercaptoethanol or chelators like EDTA/EGTA, HisPrem™ His-tag Purification Resin (Reductant/Chelator-resistant) or HisPrem™ His-tag Purification Resin (Fast Flow, Reductant/Chelator-resistant) is recommended.

This product is pre-chelated with nickel ions, giving it a blue-green color, and the gel particles have a diameter of 45-165 μm. The maximum pressure tolerance is 0.025 MPa, equivalent to approximately 5.8 psi. When performing protein purification at a fixed flow rate, the recommended flow rate is 0.5 ml/min.

This product can bind His-tagged proteins with high capacity and specificity. The maximum protein binding capacity is approximately 25-35 mg protein per mL gel (for a protein with a molecular weight of 55 kDa). The actual maximum binding capacity depends on the molecular weight of the His-tagged recombinant protein being purified, with larger molecular weights resulting in higher maximum binding capacities and smaller molecular weights resulting in lower maximum binding capacities. However, for proteins with the same molecular weight, the maximum binding capacity may vary due to differences in the protein's inherent characteristics.

This product is stored in 20% ethanol. The gel accounts for 50% of the total volume of the packaging, for example, in a total volume of 10 mL, 5 mL is gel and 5 mL is liquid. It is advisable to fully resuspend the gel before use.

Precautions

• Please do not freeze this product at temperatures of -20°C or lower.
• During the use of this product, the concentration of buffering agents such as Tris, HEPES, MOPS, etc., should not exceed 100 mM. The concentration of SDS and Sarkosyl should not exceed 0.3%. The concentration of Triton X-100, Tween-20, and NP-40 should not exceed 2%. The concentration of sodium deoxycholate and CHAPS should not exceed 1%. The concentration of histidine should not exceed 20 mM. The concentration of calcium ions should not exceed 5 mM. The concentration of sodium and magnesium ions can be as high as 2 M. The concentration of guanidine hydrochloride can be as high as 6 M. The concentration of urea can be as high as 8 M. The concentration of glycerol can be as high as 50%. This product is not compatible with reducing agents, chelators, strong acids, or strong bases. The compatibility with other unmentioned reagents can be referenced based on the aforementioned agents but requires experimental validation.
• The gel should be kept moist throughout the storage and purification process.
• We offer a range of empty columns for affinity chromatography in various sizes that are compatible with this product. If you are interested or have any specific requirements, please feel free to inquire.
• If centrifugation is not sufficient to remove insoluble substances from the protein sample, the sample solution can be filtered using a 0.45 μm filter membrane.
• After collecting the protein sample, the purification process should be completed as soon as possible, and the sample should be stored at 4°C or on ice to slow down protein degradation. To effectively inhibit protein degradation, an appropriate amount of a mixture of protease inhibitors can be added to the lysis buffer.
• If the desired purification results cannot be achieved using the conditions provided in this manual, you can try changing the concentration and/or pH of the imidazole in the wash buffer and elution buffer to achieve optimal results.
• If it is necessary to use chelators or reducing agents for the purification of the target protein, HisPrem™ His-tag Purification Resin (Reductant/Chelator-resistant) or HisPrem™ His-tag Purification Resin (Fast Flow, Reductant/Chelator-resistant) is recommended.
• This product is for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment, food, or drugs, and should not be stored in a regular residential setting.
• For your safety and health, please wear lab attire and disposable gloves when handling.

Storage

Store at 4°C. Valid for at least one year.

Only for research and not intended for treatment of humans or animals

Journals Using SBS Genetech Products                                       Universities Using SBS Genetech Products

SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory