HisPrem™ His-tag Purification Resin (Fast Flow, Denaturant-resistant)
$90.00 - $5,600.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: HPRFD-10 (for 10ml)
Cat. No.: HPRFD-100 (for 100ml)
Cat. No.: HPRFD-1k (for 1000ml)
HisPrem™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) is a His-tag protein purification medium commonly known as a nickel column, which is capable of withstanding high flow rates and pressures in denaturing conditions. This product is a novel purification medium that can tolerate high flow rates and pressures and is compatible with strong protein denaturing agents such as urea and guanidine hydrochloride. It enables simple, fast, efficient, and highly specific purification of His-tagged proteins under both non-denaturing and denaturing conditions.
Recombinant proteins containing a 6xHis-tag or more than six consecutive histidine residues are commonly referred to as His-tagged proteins. When the protein sample solution passes through HisPrem™ His-tag Purification Resin (Fast Flow, Denaturant-resistant), the histidine residues on the recombinant protein's His-tag specifically bind to the nickel ions immobilized on HisPrem™ His-tag Purification Resin (Fast Flow, Denaturant-resistant), while other proteins do not bind. After washing, the His-tagged recombinant proteins can be eluted under non-denaturing or denaturing conditions, thus separating and purifying them.
This product exhibits strong affinity, high selectivity, and high 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, through a simple, fast, and efficient purification process. The purified proteins can be used for structural and functional studies, antibody preparation, protein-protein interactions, protein-nucleic acid interactions, and other research applications.
HisPrem™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) utilizes a highly cross-linked 6% agarose gel matrix and employs optimized arm immobilization and nickel ion chelation technologies. Compared to similar products on the market, this product 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 beta-mercaptoethanol can reduce the chelated nickel ions, while divalent ion chelators such as EDTA/EGTA can cause nickel leakage. Therefore, the protein sample or purification solutions should not contain reducing agents or chelators. If protein purification is required under strong reducing agents such as DTT/beta-mercaptoethanol or chelators such as EDTA/EGTA, it is recommended to use HisPrem™ His-tag Purification Resin (Reductant/Chelator-resistant) or HisPrem™ His-tag Purification Resin (Fast Flow, Reductant/Chelator-resistant), which are tolerant to reducing agents and chelators.
This product has already been charged with nickel ions and appears blue-green in color. The gel particles have a diameter of 45-165 μm. The maximum pressure it can withstand is 0.3 MPa, equivalent to approximately 43.5 psi. When using a fixed flow rate for protein purification, the maximum flow rate is 300 cm/h.
This product can bind His-tagged proteins with high capacity and specificity. The maximum protein
binding capacity is approximately 25-35 mg protein (for proteins with a molecular weight of 55 kDa) per milliliter of gel. The actual maximum binding capacity depends on the molecular weight of the His-tagged recombinant protein to be purified. Larger proteins have a higher maximum binding capacity, while smaller proteins have a lower maximum binding capacity. However, proteins with the same molecular weight may have different maximum binding capacities due to differences in their properties.
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. Before use, it is recommended to thoroughly resuspend the gel before pipetting.
- Please do not freeze this product at temperatures equal to or below -20°C.
- During the use of this product, the concentration of buffering reagents 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 go up to 2 M. The concentration of guanidine hydrochloride can go up to 6 M. The concentration of urea can go up to 8 M. The concentration of glycerol can go up to 50%. This product is not compatible with reducing agents, chelating agents, and strong acids or bases. The compatibility with other unmentioned reagents can be referred to the aforementioned reagents, but experimental validation is required.
- The gel should always be kept moist during storage and purification processes.
- 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 does not completely 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, it is recommended to proceed with the purification process as soon as possible and always keep it at 4°C or on ice to slow down protein degradation. To effectively inhibit protein degradation, an appropriate amount of a protease inhibitor cocktail can be added to the lysis buffer.
- If the purification results are not satisfactory using the conditions provided in this manual, try changing the concentration and/or pH of the imidazole in the wash and elution buffers to achieve the optimal result.
- If the purification of the target protein requires the use of chelating agents or reducing agents, it is recommended to use HisPrem™ His-tag Purification Resin (Reductant/Chelator-resistant) or HisPrem™ His-tag Purification Resin (Fast Flow, Reductant/Chelator-resistant).
- This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in regular residential areas.
- For your safety and health, please wear lab attire and disposable gloves while handling.
Store at 4°C. Valid for at least one year.
Only for research and not intended for treatment of humans or animals
SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory