No secondary reaction is required, since the ninta complex is. The ninta resin uses nitrilotriacetic acid nta, a tetradenate chelating ligand, in a highly crosslinked 6% agarose matrix. Genscript antihis affinity resin is designed for purification of histagged protein. Ninta resin, a 6% agarose imac resin for his tag protein. Qiagen ninta agarose, 25ml, 45 to 165m bead, up to 50mgml binding capacity, cell lysate start material, nickelcharged resin, 2. High quality ninta products 80 mgml protein cube biotech. Prepacked 1 ml and 5 ml columns with precharged nickelnta agarose resin. The qiaexpressionist 062003 3 contents kit contents 7 storage conditions 8 technical assistance 9 product use limitations 9 product warranty and satisfaction guarantee 9 safety information 10 introduction and general guidelines 11 the qiaexpress system 15 qiaexpress pqe vectors 15 qiaexpress pqetrisystem vector for expression in li, mammalian and. Ninta column preparation resuspend ninta agarose slurry in a bottle container. The purification technique is based on the strong affinity of. 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. It is composed of agarose and conjugated the tm antihis monoclonal antibody. Retrieve the dialyzed sample from 4c and place it on ice.
Qiagen ninta agarose, 100ml, 45 to 165m bead, up to 50mgml binding capacity, cell lysate start material, nickelcharged resin, 2. In this protocol binding is performed in batch mode in contrast to oncolumn binding, since it is. Cgi114 protein concentrations were measured by absorbance at 280 nm mw 26,275. Your new discounted price will be automatically recalculated. 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. Qiagen ninta spin kit, 16 to 24m bead, up to 300gspin column binding capacity, cell lysate start material, automated processing, macroporus silica support, up to 95% homogeneity in one step, for fast, smallscale. This resin consists of crosslinked agarose derivatized with nitrilotriacetic acid nta and provides good properties working in native or denaturing conditions. 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 nickelnitrilo triacetic acid ninta coupled to sepharose cl6b from qiagen offers a quick and efficient method of protein purification from small and large scale preparations obtained through variable expression sources like e. Nintaatto conjugates can be directly applied either to an sdspage gel or western blot membrane for fluorescence imaging, and have been successfully used in living cells. Ninta agarose uses nta which represents the most commonly used chelating ligand in imac. Ni nta agarose purification of 6xhistagged proteins from e. Highlyefficient purification of native polyhistidinetagged. 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 ninta spin kit, 16 to 24m bead, up to 300gspin column binding capacity, cell lysate start material, automated processing, macroporus silica support, up to 95% homogeneity in one step, for fast, smallscale purification of 6xhis tagged proteins, ideal for proteomics application, includes 50 ninta spin columns, reagents, buffers. The protein yield in 4 elutions e1e4, cube was 80 mgml, compared to 65 and 48 mgml obtained with the alternative resins e1e4, competitor g, competitor q. Highlyefficient purification of native polyhistidine. The 6xhisninta system allows rapid and efficient affinity purification of recombinant proteins from virtually any expression system. Ni affinity column can effectively purify extract even just one column ni nta agarose in small column bulk materials with very competitive prices. Onestep purification can be performed under both native and denaturing conditions. Protocols and tips for purification under both native and denaturing conditions are provided, as well as a rapid spin procedure for protein minipreps. 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 applications. Download the kingfisher duo bindit software protocol. Place 50ul beads 100ul suspension of ninta agarose beads in 1. 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. Sep 06, 2007 the nickelnitrilo triacetic acid ni nta coupled to sepharose cl6b from qiagen offers a quick and efficient method of protein purification from small and large scale preparations obtained through variable expression sources like e. All reagents are guaranteed stable for 6 months when properly stored.
Dear all, i am using ninta agarose qiagen for purification of histagged proteins by gravityflow chromatography. The reuse of ninta resin depends on the nature of the sample and should only be performed with identical recombinant proteins. Ac501 purification histagged proteins nickel nta agarose. Nitrilotriacetate nickel an overview sciencedirect topics. Basel, switzerland, who have purified more than 100 different proteins on ninta resin, we recommend a maximum of 5 runs per column. Ni nta technology 18 ni nta chromatographic materials 19 cloning 21 choosing a qiaexpress construct 21 intended use of recombinant proteins and pqe vector choice 22 protein size 22 codon usage 23 internal start sites 23 inefficient translation 23 secretion 23 cloning procedures and vector maps 24 procedure for direct cloning of pcr fragments. Qiagen ni nta spin kit, 16 to 24m bead, up to 300gspin column binding capacity, cell lysate start material, automated processing, macroporus silica support, up to 95% homogeneity in one step, for fast, smallscale purification of 6xhis tagged. Ni nta column preparation resuspend ni nta agarose slurry in a bottle container. For purification of histagged proteins by gravityflow chromatography.
Oem manufacturer in china, for ni nta agarose, deae agarose. How imidazole can be washed off the ninta agarose beads. Histagged protein purification, protein and peptide purification, protein sample preparation and protein purification, proteins, expression, isolation and analysis. 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. 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. Step 2a batch purification of histagged proteins from e. Let the resin settle by gravity and gently aspirate the supernatant.
Add 6 ml of sterile distilled water and resuspend resin. Due to the high affinity, ninta magnetic beads can be used for capturing 6x histagged proteins. Nitrilotriacetic acid is commercially available as the free acid and as the sodium salt. Ninta buffer kit 708993 bugbuster ninta hisbind purification kit 7075 description ninta hisbind resins store ninta hisbind resin and ninta hisbind superflow at 4c. Basel, switzerland, who have purified more than 100 different proteins on ni nta resin, we recommend a maximum of 5 runs per column. The ni nta resin can be used to purify 6x his tagged proteins under native and 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 applications. Jul 28, 2003 all chemicals are purchased from merck except for nintaagarose qiagen, glutathioneagarose sigma and coomassie brilliant blue g 250 serva. Ninta purification system thermo fisher scientific. For manual or automated purification of histagged proteins pdf 111kb.
Nickel nta agarose beads are provided readytouse for rapid purification of histagged proteins under native or denaturing conditions. Ni nta column preparation resuspend ninta agarose slurry in a bottle container. Gfp was spiked into li lysates in defined quantities 10, 5, 2. Ninta spin kit 50 314 from qiagen sample to insight. Nitrilotriacetic acid nta is the aminopolycarboxylic acid with the formula n ch 2 co 2 h 3. Oem manufacturer in china, for ni nta agarose, deae. Ordering information qiagen promotion march 2017 until march 24, 2017 you can save 2 on selected assay and sample prepration products when you order 4% through your eprocurement system. The spin columns are supplied with a resin bed volume of 0. All chemicals are purchased from merck except for nintaagarose qiagen, glutathioneagarose sigma and coomassie brilliant blue g 250 serva. Ni nta agarose, deae agarose, deae cellulose de52, cm. Gbiosciences ninta magnetic beads are 3m beads designed for the rapid purification of x histagged proteins. Qiagen ninta hissorb plate, 96well, transparent, 5 nintacoated, 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 6xhis.
This resin can recover histagged proteins from a variety of expression systems such as baculovirus, yeast, mammalian and bacterial cells. The metal ions are immobilized on chromatographic matrices by a chelating ligand, most commonly nitrilotriacetic acid nta or iminodiacetic acid ida. Pressure, manualautomated processing, large scale, sepharose cl6b matrix, 100g to 100mg yield, 6xhis. Given that this agarose is remarkably expensive i would like to reuse it. Ni nta agarose is a nickelcharged affinity resin that can be used to purify recombinant proteins containing a polyhistidine 6xhis sequence. They are not to be used for human diagnostic or drug purposes. Ninta magnetic beads have nitrilotriacetic acid nta groups with charged nickel covalently bound to the surface dextran of the beads. Ni nta agarose beads are widely used for protein purification due to its high affinity and selectivity for recombinant fusion proteins that are tagged with six tandem histidine residues. Onestep purification of recombinant proteins with the. Excellent protein recovery with purecube 100 ninta agarose. Ninta column purification skip step 4 procedures contained here if purifying homodimer.
Ni nta is designed for high quality purification of 6xhistagged recombinant proteins from bacteria, insect and mammalian cells. Over 20% more yield obtained with purecube ninta agarose. Small scale histag purification under nature conditions. The solvent was removed under reduced pressure to give the 53,4. Excellent protein recovery with purecube 100 ni nta agarose. The ninta resin can be used to purify 6x his tagged proteins under native and denaturing conditions. Purification of histagged proteins by imac is based on the affinity of histidine residues for immobilized metal ions e. Ni nta magnetic agarose beads are used for smallscale affinity purification as well as highthroughput screening of recombinant histagged proteins.
Ni nta agarose uses nta which represents the most commonly used chelating ligand in imac. During this step, the histagged target protein is purified under native conditions from the cleared e. Ninta spin kit handbook 012008 5 product use limitations qiaexpress products are developed, designed, and sold for research use. Synthesis of 53,4bisbenzyloxyphenethylcarbamoylbenzene1,3benzylntalysine the diacid 2 0. These results are suggestive of similar binding affinity of mono and bisniiintamodified nanoparticles, probably a result of high surface coverage of.
Ni nta agarose, ni nta superflow manufacturersupplier. For fast, smallscale purification of histagged proteins, includes spin columns, reagents, buffers, and control plasmid. Qiagen ni nta agarose, 100ml, 45 to 165m bead, up to 50mgml binding capacity, cell lysate start material, nickelcharged resin, 2. Dear all, i am using ni nta agarose qiagen for purification of histagged proteins by gravityflow chromatography. Ninta agarose and purification columns have the following specifications. Histagged protein in the crude samples can bind to the antihis monoclonal antibodies coupled with the agarose, and the nonspecific proteins can be eliminated by washing. Proteins bound to the resin can be eluted with low ph buffer or competition with imidazole or histidine. Prepare a 15 ml ninta agarose resin column in a 4 c chromatography system. Ninta agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a his tag. The ni nta resin uses nitrilotriacetic acid nta, a tetradenate chelating ligand, in a highly crosslinked 6% agarose matrix. Ninta agarose is a nickelcharged affinity resin that can be used to purify recombinant proteins containing a polyhistidine 6xhis sequence. This resin can recover histagged proteins from a variety of expression systems such as.
Ordering information qiagen promotion march 2017 until march 24. The polyhistidine tag is the most popular affinity tag and typically consists of six consecutive histidine residues 6xhis. Invitrogen novex ninta purification system 6 purifications. Ninta agarose beads are widely used for protein purification due to its high affinity and selectivity for recombinant fusion proteins that are tagged with six tandem histidine residues. Ninta is designed for high quality purification of 6xhistagged recombinant proteins from bacteria, insect and mammalian cells. The reuse of ni nta resin depends on the nature of the sample and should only be performed with identical recombinant proteins. Recovered gfp was quantified by absorption at 488 nm. Qiagen ni nta agarose, 25ml, 45 to 165m bead, up to 50mgml binding capacity, cell lysate start material, nickelcharged resin, 2. Qiagen ni nta spin kit, 16 to 24m bead, up to 300gspin column binding capacity, cell lysate start material, automated processing, macroporus silica support, up to 95% homogeneity in one step, for fast, smallscale purification of 6xhis tagged proteins, ideal for proteomics application, includes 50 ni nta spin columns, reagents, buffers.
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