Utilizes ninta nickelchelating resin and is supplied with native and denaturing buffers for efficient purification of recombinant proteins under different conditions histagged protein. 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. Ninta superflow cartridges or qiagen products in general, please. Ni nta agarose beads are widely used for protein purification due to its high affinity and selectivity for recombinant fusion. They are used for immobilizing and purifying recombinant. Easyxpress protein synthesis handbook harvard university. With the 5 ml bed filled with ninta agarose from qiagen, the total price of a.
Nov 22, 2015 ninta superflow cartridge handbook for manual or fplc purification of. Qiaexpress detection and assay handbook third edition. Reuse of qiagen ni nta columns after dry out what are your. Ninta magnetic agarose beads are magnetic particles coated with ninta agarose affinity purification matrix. 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. 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. Ni nta agarose uses nta which represents the most commonly used chelating ligand in imac. 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. The ni nta purification system is a complete system that includes purification buffers and resin for purifying proteins under native, denaturing, or hybrid conditions. High dynamic binding capacity of purecube 100 ni nta agarose. Highlyefficient purification of native polyhistidinetagged proteins by multivalent nta modified magnetic nanoparticles. Then go to the qiagen web shop, your organizations purchasing system, your sales rep, or by email and order a minimum of three kits. A handbook for highlevel expression and purification of 6xhis tagged. Ninta affinity purification of histagged proteins is a bindwashelute procedure that can be performed under native or denaturing conditions.
How imidazole can be washed off the ninta agarose beads. 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. The ninta purification system is a complete system that includes purification buffers and resin for purifying proteins under native, denaturing, or hybrid conditions. Genscript antihis affinity resin is designed for purification of histagged protein. Purecube ni nta agarose is robust against oxidation and regenerable. Ninta magnetic agarose beads under native conditions. Ninta spin kit handbook 012008 5 product use limitations qiaexpress products are developed, designed, and sold for research use. Please read the ninta superflow 96 biorobot kit handbook carefully before beginning any of these procedures. Ninta superflow cartridge handbook for manual or fplc purification of. Qiagen ninta hissorb plate, 96well, transparent, 5 nintacoated, histidine residued epitope detected.
Ninta agarose is an affinity chromatography matrix for purifying. It is composed of agarose and conjugated the tm antihis monoclonal antibody. Ninta resin, a 6% agarose imac resin for his tag protein. Highlyefficient purification of native polyhistidinetagged. 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. Print bookmark share more for manual or automated purification of histagged proteins. The ni nta resin can be used to purify 6x his tagged proteins under native and denaturing conditions. Ninta spin handbook the wolfson centre for applied structural. Add 6 ml of sterile distilled water and resuspend resin. Most manufacturers recommend using 48 times before regeneration. Original manufacturer offers bulk volume of deae agarose, ni nta. 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. Purecube ninta agarose is robust against oxidation and regenerable.
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. Ninta agarose is a nickelcharged affinity resin that can be used to purify recombinant proteins containing a polyhistidine 6xhis sequence. 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. Handbook 1 ni nta hissorb strips 24 ni nta hissorb plates 5.
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. Highlyefficient purification of native polyhistidine. Proteins bound to the resin can be eluted with low ph buffer or competition with. Ninta superflow is comprised of ninta coupled to superflow resin. The ni nta resin uses nitrilotriacetic acid nta, a tetradenate chelating ligand, in a highly crosslinked 6% agarose matrix. Hifliq ninta fplc columns are supplied prepacked and ready to use with precharged nickelnta agarose resin for affinity purification of polyhistidine. Oem manufacturer in china, for ni nta agarose, deae. In my lab we package ninta agarose into pd10 or nap10 columns, and used portions are pooled and stored at 4c in 20%.
Reuse of qiagen ni nta columns after dry out what are your thoughts. Ninta agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a his tag. For manual or automated purification of histagged proteins pdf 111kb. Proteins bound to the resin may be eluted with either low ph buffer or by competition with imidazole or histidine. Material required the imidazole concentrations of elution buffers 1 and 2 must be optimized for each protein. 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. 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.
Let the resin settle by gravity and gently aspirate the supernatant. Ni nta superflow 500 ml 500 ml nickelcharged resin max. Ni nta column preparation resuspend ni nta agarose slurry in a bottle container. Ninta superflow biorobot handbook supplied with the columns or contact one of the qiagen. Ni affinity column can effectively purify extract even just one column. Material required the imidazole concentrations of elution buffers 1 and 2 must be optimized for each. Here, protocols for purification of histagged proteins under native, as well as under denaturing conditions, are given. Ninta superflow biorobot handbook supplied with the columns or contact.
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. Histidine residues in the his tag bind to the vacant positions in the coordination. Ni nta agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a his tag. Histagged protein purification, protein and peptide purification, protein sample preparation and protein purification, proteins, expression, isolation and analysis. The ni nta agarose contained in this product is manufactured by qiagen.
Proteins bound to the resin can be eluted with low ph buffer or competition with imidazole or histidine. 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. If the resin changes from light blue to brownishgray the nickel has been. Dear all, i am using ninta agarose qiagen for purification of histagged proteins by gravityflow chromatography. Manual purification of 6xhistagged proteins from e. This protein purification system is based on the remarkable selectivity of our unique. This product is not intended for the diagnosis, prevention, or treatment of a disease. Ni nta agarose, deae agarose, deae cellulose de52, cm.
The polyhistidine tag is the most popular affinity tag and typically consists of six consecutive histidine residues 6xhis. Gfp was spiked into li lysates and purified on a 1 ml purecube cartridge filled with purecube 100 ninta. Qiagen ninta agarose, 25ml, 45 to 165m bead, up to 50mgml binding capacity. 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. Ninta agarose is an affinity chromatography matrix for purifying recombinant.
How imidazole can be washed off the ninta agarose beads reuse of ninta agarose beads for his protein reply. Ninta is designed for high quality purification of 6xhistagged recombinant proteins from bacteria, insect and mammalian cells. Technical service departments or local distributors listed on. Ninta magnetic agarose beads are used for smallscale affinity purification as well as highthroughput screening of recombinant histagged proteins. The nickel nitrilotriacetic acid nta is a qiagenpatented resin which offers affinity purification of 6xhistagged proteins expressed as recombinant proteins from sources. They are not to be used for human diagnostic or drug purposes.
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. Ni nta is designed for high quality purification of 6xhistagged recombinant proteins from bacteria, insect and mammalian cells. Ac501 purification histagged proteins nickel nta agarose. Ninta magnetic agarose beads 16 detection of 6xhistagged proteins 17 immobilization onto membrane western, dot, or colony blotting 20. Ninta magnetic beads have nitrilotriacetic acid nta groups with. Ninta superflow cartridge handbook the wolfson centre for. 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. Highlyefficient purification of native polyhistidinetagged proteins by multivalent ntamodified magnetic nanoparticles. Check that the resin is contained in the narrow part of the column body before. Ninta purification system thermo fisher scientific. Qiagen ni nta agarose, 25ml, 45 to 165m bead, up to 50mgml binding capacity, cell lysate start material, nickelcharged resin, 2. 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. Qiagen will replace it free of charge or refund the purchase.
Ninta superflow cartridges or qiagen products in general, please do not. Ni nta magnetic agarose beads are used for smallscale affinity purification as well as highthroughput screening of recombinant histagged proteins. Qiagen ninta agarose, 25ml, 45 to 165m bead, up to. A handbook for highlevel expression and purification of 6xhistagged proteins. Ni nta affinity purification of histagged proteins is a bindwashelute procedure that can be performed under native or denaturing conditions. Ninta agarose and purification columns have the following specifications.
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. So, i would like to know how to reuse the ninta column qiagen with. The ninta resin can be used to purify 6x his tagged proteins under native and denaturing conditions. For purification of histagged proteins by gravityflow chromatography.
Nickel columns are used for immobilized metal affinity chromatography imac for the purification of recombinant proteins with a polyhistidine tag on either terminus. 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. Ni nta agarose 500ml nta agarose purification from left to right 1. Ninta column preparation resuspend ninta agarose slurry in a bottle container. High dynamic binding capacity of purecube 100 ninta agarose. Oem manufacturer in china, for ni nta agarose, deae agarose. Purification of in vitro synthesized proteins 44 using ninta magnetic agarose beads 44 using ninta magnetic. Purecube ninta agarose was exposed to 5mm dtt for 1 h a. Reagent vendor ninta agarose qiagen 1 ml column with luer lock on both ends mobitec 10 ml luer lock syringe merck eurolab buffer composition. At qiagen we pride ourselves on the quality and availability of our.
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