Protein purification is used to isolate recombinant proteins and antibodies fused with Flag tag, GST tag or His tag in an efficient manner, among which His-tag protein purification has been extensively used for more than 30 years. The principle of His-tag protein purification is that the histidine (His) residue carries an imidazole group which can selectively bind to metal ions such as Ca2+, Mg2+, Ni2+ and Co2+, of which Ni2+ is the most widely used in affinity purification. There are many kinds of ligands used to prepare metal chelating media, and the commonly used ones are iminodiacetic acid (IDA), nitrilotriacetic acid (NTA), and tris(carboxymethyl)ethylenediamine (TED).
As we all know, protein purification is a very hard task, and for slightly longer gravity flow columns, the whole purification process will be extremely long. To solve this problem, Beyotime has developed new His-tag protein purification media including BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) (P2221) and BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236) that have high flow rate and can tolerate high pressures.
BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant)
Beyotime's BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) (P2221) is a new purification resin that is compatible with reductants and chelators with high flow rates and high pressure tolerance, and enables simple, rapid, efficient and highly specific purification of His-tagged proteins. The histidine residues of the His-tag of recombinant proteins specifically bind to the nickel ion in the BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant), while other proteins cannot. After washing, the purified His-tagged recombinant protein can be obtained by elution under native conditions.
Typical Figures:
Figure 1. The purification result of His-tagged recombinant proteins using Beyotime's BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) (P2221). M. marker; CL. cell lysate; FT. flow through; W1-W3. wash 1-3; E1-E4. eluate 1-4.
BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236)
Beyotime's BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236) is a purification resin that is compatible with denaturants (e.g., urea and guanidine hydrochloride) with high flow rates and high pressure tolerance, and enables simple, rapid, efficient and highly specific purification of His-tagged proteins under either native or denaturing conditions.
Typical Figures:
Protein purification under native conditions
Figure 2. The purification result of His-tagged recombinant proteins using Beyotime's BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236) under native conditions. M. marker (Beyotime, P0076); CL. cell lysate; FT. flow through; W1-W3. wash 1-3 (native lysis buffer); W4-W8. wash 4-8 (native lysis buffer with 20mM imidazole); E1-E4. eluate 1-4 (native lysis buffer with 250mM imidazole).
Protein purification under denaturing conditions with 8M urea
Figure 3. The purification result of His-tagged recombinant proteins using Beyotime's BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236) under denaturing conditions with 8M urea. All solutions used contained 8M urea. M. marker (Beyotime, P0076); CL. cell lysate; FT. flow through; W1-W3. wash 1-3 (denaturing lysis buffer); W4-W8. wash 4-8 (denaturing lysis buffer with 20mM imidazole); E1-E4. eluate 1-4 (denaturing lysis buffer with 250mM imidazole).
Protein purification under denaturing conditions with 6M guanidine hydrochloride
Figure 4. The purification result of His-tagged recombinant proteins using Beyotime's BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236) under denaturing conditions with 6M guanidine hydrochloride. All the solutions used contained 6M guanidine hydrochloride, and the guanidine hydrochloride in each sample was removed by dialysis before electrophoresis. M. marker (Beyotime, P0076); CL. cell lysate; FT. flow through; W1-W3. wash 1-3 (denaturing lysis buffer); W4-W8. wash 4-8 (denaturing lysis buffer with 20mM imidazole); E1-E4. eluate 1-4 (denaturing lysis buffer with 250mM imidazole).
Product Features:
▶ Highly resistant to pressure: These products are more tolerant of high flow rates and pressures than previously introduced BeyoGold™ His-tag Purification Resin (Reductant&Chelator-resistant) (P2210/P2218/P2220) and BeyoGold™ His-tag Purification Resin (Denaturant-resistant) (P2233). It can tolerate pressure up to 0.3 MPa (~43.5 psi) and a maximum flow rate of 300 cm/h.
▶ High purity of His-tagged recombinant protein can be obtained: With high affinity for His-tagged recombinant proteins, high selectivity and high binding capacity, high purity of His-tagged recombinant proteins can be obtained by using these products.
▶ High amount of His-tagged recombinant protein can be obtained: BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) has a maximum protein binding capacity of 20-30 mg protein/ml gel. BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) has a maximum protein binding capacity of approximately 25-35 mg protein (55 kD protein)/ml gel.
▶ Reduced non-specific protein binding: These products are based on a highly cross-linked 6% agarose gel (essentially the same performance as Sepharose 6 Fast Flow) with optimizations of splice arms and nickel ion chelation. This new resin enables significantly reduced non-specific protein binding, higher stress tolerance and more stable nickel ion chelation compared to other commonly used Ni-NTA agarose or similar products.
▶ Very stable nickel ion chelation: Compared with similar products, these two products do not need to reload nickel ions for reuse because nickel ions are basically not lost.
▶ Wide applications of purified proteins: The purified proteins can be used for structural and functional studies, antibody preparation, protein-protein interactions, and protein-nucleic acid interactions.
Tips：
In addition to the features mentioned above, BeyoGold™ His-tag Purification Resin (Fast Flow, Reductant&Chelator-resistant) (P2221) is highly resistant to reducing agents such as DTT and chelators such as EDTA, offering the possibility of purification of protein samples requiring the presence of reducing or chelating agents.
BeyoGold™ His-tag Purification Resin (Fast Flow, Denaturant-resistant) (P2236) can tolerate 8M urea or 6M guanidine hydrochloride and can be used for the purification of His-tagged proteins under either native or denaturing conditions.
Product Information: