Manual purification of 6xhistagged proteins from e. For purification of histagged proteins by gravityflow chromatography. Ni nta magnetic agarose beads are used for smallscale affinity purification as well as highthroughput screening of recombinant histagged proteins. How imidazole can be washed off the ninta agarose beads reuse of ninta agarose beads for his protein reply. Ni nta agarose 500ml nta agarose purification from left to right 1. Hifliq ninta fplc columns are supplied prepacked and ready to use with precharged nickelnta agarose resin for affinity purification of polyhistidine. Ninta magnetic agarose beads under native conditions. Highlyefficient purification of native polyhistidinetagged proteins by multivalent nta modified magnetic nanoparticles. Ni nta agarose is a nickelcharged affinity resin that can be used to purify recombinant proteins containing a polyhistidine 6xhis sequence. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Technical service departments or local distributors listed on. Ninta superflow cartridge handbook 032007 7 introduction qiagen ninta superflow cartridges are prefilled with 1 ml or 5 ml ninta superflow and are ready to use for purification of 6xhistagged proteins using a syringe, peristaltic pump, or liquid chromatography system such as the aktadesign or fplc system. They are used for immobilizing and purifying recombinant. Ninta superflow cartridge handbook the wolfson centre for. The ninta resin can be used to purify 6x his tagged proteins under native and denaturing conditions. Let the resin settle by gravity and gently aspirate the supernatant. Ninta agarose is an affinity chromatography matrix for purifying. Add 6 ml of sterile distilled water and resuspend resin. Ninta superflow cartridges or qiagen products in general, please do not. The polyhistidine tag is the most popular affinity tag and typically consists of six consecutive histidine residues 6xhis. Ninta agarose consists of the chelating ligand nitrilotriacetic acid nta immobilized on 6 % crosslinked agarose beads that are suitable for batch binding, gravity flow, and fplc columns. Gfp was spiked into li lysates and purified on a 1 ml purecube cartridge filled with purecube 100 ni nta agarose at flow rates from 1 to 7 mlmin, and in a gravity flow batch procedure.
The ni nta agarose contained in this product is manufactured by qiagen. Dear all, i am using ninta agarose qiagen for purification of histagged proteins by gravityflow chromatography. Ninta magnetic beads have nitrilotriacetic acid nta groups with. Ninta agarose and purification columns have the following specifications. Highlyefficient purification of native polyhistidinetagged proteins by multivalent ntamodified magnetic nanoparticles. Material required the imidazole concentrations of elution buffers 1 and 2 must be optimized for each. How imidazole can be washed off the ninta agarose beads. Ninta superflow cartridge handbook for manual or fplc purification of.
At qiagen we pride ourselves on the quality and availability of our. Histagged protein purification, protein and peptide purification, protein sample preparation and protein purification, proteins, expression, isolation and analysis. If the resin changes from light blue to brownishgray the nickel has been lost or changed its oxidations state and will no longer bind his tagged proteins. Qiaexpress detection and assay handbook third edition. Ni nta agarose, deae agarose, deae cellulose de52, cm. High dynamic binding capacity of purecube 100 ni nta agarose. Print bookmark share more for manual or automated purification of histagged proteins.
Qiagen ninta agarose, 25ml, 45 to 165m bead, up to 50mgml binding capacity. Ninta superflow biorobot handbook supplied with the columns or contact one of the qiagen. The nickel nitrilotriacetic acid nta is a qiagenpatented resin which offers affinity purification of 6xhistagged proteins expressed as recombinant proteins from sources. Nickel columns are used for immobilized metal affinity chromatography imac for the purification of recombinant proteins with a polyhistidine tag on either terminus. Qiagen distributors please see the last page for contact information for your local qiagen distributor. Ninta resin for recombinant proteins carrying a small affinity tag consisting of. Nickel nta agarose beads are provided readytouse for rapid purification of histagged proteins under native or denaturing conditions. Check that the resin is contained in the narrow part of the column body before. Oem manufacturer in china, for ni nta agarose, deae agarose. Reusing ni nta resin questions so i havent been able to find this in the manuals but, we do a lot of routine protein purification and i was never trained to do anything with the resin other than wash it excessively with water before storing it in 20% ethanol.
Ninta superflow cartridges or qiagen products in general, please. Ninta magnetic agarose beads 16 detection of 6xhistagged proteins 17 immobilization onto membrane western, dot, or colony blotting 20. Please read the ninta superflow 96 biorobot kit handbook carefully before beginning any of these procedures. Ni nta agarose consists of the chelating ligand nitrilotriacetic acid nta immobilized on 6 % crosslinked agarose beads that are suitable for batch binding, gravity flow, and fplc columns. Ni nta affinity purification of histagged proteins is a bindwashelute procedure that can be performed under native or denaturing conditions. Qiagen ninta hissorb plate, 96well, transparent, 5 nintacoated, histidine residued epitope detected. This offer is valid until december 31, 2019 and only when order is placed online through qiagen web shop, your organizations purchasing system, your sales rep, or by email using the cyber19 promo code. Original manufacturer offers bulk volume of deae agarose, ni nta. Purification of in vitro synthesized proteins 44 using ninta magnetic agarose beads 44 using ninta magnetic. Here, protocols for purification of histagged proteins under native, as well as under denaturing conditions, are given. Reuse of qiagen ni nta columns after dry out what are your thoughts. It is composed of agarose and conjugated the tm antihis monoclonal antibody.
Ni affinity column can effectively purify extract even just one column. Proteins bound to the resin may be eluted with either low ph buffer or by competition with imidazole or histidine. So, i would like to know how to reuse the ninta column qiagen with. Ninta agarose 10 ml 10 ml disposable columns, 5 ml bedvolume 5 5 disposable columns, 1 ml bedvolume 5 5 frits for 5 ml 10 10 disposable columns. High dynamic binding capacity of purecube 100 ninta agarose. Proteins bound to the resin can be eluted with low ph buffer or competition with imidazole or histidine. Oem manufacturer in china, for ni nta agarose, deae. Qiagen ni nta agarose, 25ml, 45 to 165m bead, up to 50mgml binding capacity, cell lysate start material, nickelcharged resin, 2.
If the resin changes from light blue to brownishgray the nickel has been. A handbook for highlevel expression and purification of 6xhis tagged. Histidine residues in the his tag bind to the vacant positions in the coordination sphere of the immobilized nickel ions with high specificity and affinity. Ni nta agarose beads are widely used for protein purification due to its high affinity and selectivity for recombinant fusion. Ninta superflow biorobot handbook supplied with the columns or contact. Most manufacturers recommend using 48 times before regeneration.
The ni nta resin uses nitrilotriacetic acid nta, a tetradenate chelating ligand, in a highly crosslinked 6% agarose matrix. Ninta magnetic agarose beads are magnetic particles coated with ninta agarose affinity purification matrix. Ninta agarose is an affinity chromatography matrix for purifying recombinant. Reuse of qiagen ni nta columns after dry out what are your. Purecube ni nta agarose was exposed to 5mm dtt for 1 h a. How imidazole can be washed off the ninta agarose beads reuse of ninta agarose beads for his protein jan122005 can someone guide me on how to get rid of imidazole adsorbed to. Nov 22, 2015 ninta superflow cartridge handbook for manual or fplc purification of. The ni nta purification system is a complete system that includes purification buffers and resin for purifying proteins under native, denaturing, or hybrid conditions. It occurred to me after reading the handbook that a you shouldnt reuse the resin more than 5 times b you need to rinse with 0. Hifliq ninta fplc columns user guide protein arks hifliq ninta fplc columns designed for rapid onestep purification, and ideal for preparative purification and contaminant removal. With the 5 ml bed filled with ninta agarose from qiagen, the total price of a. Ninta affinity purification of histagged proteins is a bindwashelute procedure that can be performed under native or denaturing conditions.
Ni nta agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a his tag. Highlyefficient purification of native polyhistidinetagged. Ac501 purification histagged proteins nickel nta agarose. Ninta agarose is a nickelcharged affinity resin that can be used to purify recombinant proteins containing a polyhistidine 6xhis sequence. Ninta uses the chelating ligand nitrilotriacetic acid nta coupled to a crosslinked 6% agarose resin that is suitable for use in batch and gravity flow applications. This protein purification system is based on the remarkable selectivity of our unique. Ninta spin kit handbook 012008 5 product use limitations qiaexpress products are developed, designed, and sold for research use. Reagent vendor ninta agarose qiagen 1 ml column with luer lock on both ends mobitec 10 ml luer lock syringe merck eurolab buffer composition. The ninta purification system is a complete system that includes purification buffers and resin for purifying proteins under native, denaturing, or hybrid conditions. Original manufacturer offers bulk volume of deae agarose, ni nta agarose, citacron blue agarose, cobalt nta agarose, deae cellulose de52, magnetic agarose beads and plain agarose beads. They are not to be used for human diagnostic or drug purposes. Ninta agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a his tag.
Ni nta is designed for high quality purification of 6xhistagged recombinant proteins from bacteria, insect and mammalian cells. Genscript antihis affinity resin is designed for purification of histagged protein. Ninta spin handbook the wolfson centre for applied structural. Ninta resin, a 6% agarose imac resin for his tag protein. Purecube ninta agarose was exposed to 5mm dtt for 1 h a. Ninta purification system thermo fisher scientific. Easyxpress protein synthesis handbook harvard university. Ninta column preparation resuspend ninta agarose slurry in a bottle container. Ninta magnetic agarose beads are supplied as a 5% vv suspension with a binding capacity of 300 g protein per ml of suspension for 6xhis. Ni nta agarose uses nta which represents the most commonly used chelating ligand in imac. Then go to the qiagen web shop, your organizations purchasing system, your sales rep, or by email and order a minimum of three kits. Ni nta superflow 500 ml 500 ml nickelcharged resin max. Ninta superflow is comprised of ninta coupled to superflow resin. The ni nta resin can be used to purify 6x his tagged proteins under native and denaturing conditions.
Material required the imidazole concentrations of elution buffers 1 and 2 must be optimized for each protein. Purecube ninta agarose is robust against oxidation and regenerable. Qiagen ninta agarose, 25ml, 45 to 165m bead, up to. Gfp was spiked into li lysates and purified on a 1 ml purecube cartridge filled with purecube 100 ninta. Equilibrate the ninta superflow resin by adding 600 l buffer npi10 to each well and apply a vacuum for approximately 2 min or until the buffer has been. Utilizes ninta nickelchelating resin and is supplied with native and denaturing buffers for efficient purification of recombinant proteins under different conditions histagged protein. Purification of in vitro synthesized proteins 44 using ni nta magnetic agarose beads 44 using ni nta magnetic agarose beads in 96well plates 45 using ni nta superflow under native conditions 47 using streptactin magnetic beads 48 using streptactin superflow 49.
Ni nta column preparation resuspend ni nta agarose slurry in a bottle container. Handbook 1 ni nta hissorb strips 24 ni nta hissorb plates 5. In my lab we package ninta agarose into pd10 or nap10 columns, and used portions are pooled and stored at 4c in 20%. Ni nta uses the chelating ligand nitrilotriacetic acid nta coupled to a crosslinked 6% agarose resin that is suitable for use in batch and gravity flow applications. Qiagen ni nta hissorb plate, 96well, transparent, 5 ni nta coated, histidine residued epitope detected, plate based format, manual instrument, ligand on plate, multiwellbased assay system, enhanced signaltonoise ratio and reproducibility, reduced optimization, for assays using 6xhistagged biomolecules. Highlyefficient purification of native polyhistidine. A handbook for highlevel expression and purification of 6xhistagged proteins. Ninta magnetic agarose beads are used for smallscale affinity purification as well as highthroughput screening of recombinant histagged proteins. Proteins bound to the resin can be eluted with low ph buffer or competition with. Histidine residues in the his tag bind to the vacant positions in the coordination. Purecube ni nta agarose is robust against oxidation and regenerable. Ninta superflow cartridge handbook 032007 7 introduction qiagen ninta superflow cartridges are prefilled with 1 ml or 5 ml ninta superflow and are ready to use for.
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