Protein PurificationCrystallization9Fractionation & Depletion10Tagged Protein Expression, Purification, Detection37Ni-NTA AgarosePrint For purification of His-tagged proteins by gravity-flow chromatography One-step purification from crude lysate to >95% pure proteinHigh binding affinity and high capacity Choice of purification under native or denaturing conditions Precharged, ready-to-use matrices for any scale of purification Automated purification and assay protocols Ni-NTA Agarose is an affinity chromatography matrix for purifying recombinant proteins carrying a His tag. 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. Cleared cell lysates are loaded onto the matrices. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, His-tagged proteins are eluted in buffer under native or denaturing conditions. Buy ProductsProduct DetailsSpecificationsProduct ResourcesBuy ProductsCat No./ID:30210Ni-NTA Agarose (25 ml)$323.00Order Product25 ml nickel-charged resin (max. pressure: 2.8 psi)Cat No./ID:30230Ni-NTA Agarose (100 ml)$1,114.00Order Product100 ml nickel-charged resin (max. pressure: 2.8 psi)Cat No./ID:30250Ni-NTA Agarose (500 ml)$4,818.00Order Product500 ml nickel-charged resin (max. pressure: 2.8 psi)The Ni-NTA Agarose is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.Product Details One-step purification under native conditions.One-Step Purification under Native Conditions. Human serum response factor (SRF) was expressed in HeLa cells carrying a vaccinia virus vector and purified using Ni-NTA Agarose with different imidazole concentrations in the wash and elution steps. Proteins were visualized by silver staining. CL: cell lysate; FT: flow-through; W1: 0.8 mM wash; W2 & W3: 8 mM wash; W4 & W5: 40 mM wash; E1 & E2: 80 mM elution. (Data kindly provided by H. Stunnenberg, EMBL, Heidelberg, Germany.)Protein purification with the Ni-NTA protein purification system.Performance Ni-NTA Agarose provides Ni-NTA coupled to a Sepharose CL-6B support and offers high binding capacity and minimal nonspecific binding (see figure One-step purification under native conditions). This material has excellent handling properties for most scales of batch and column purification. Ni-NTA Agarose is available separately or as a component of QIAexpress Kits, which are complete kits for efficient expression and purification of His-tagged proteins from E. coli . Principle The QIAexpress Ni-NTA Protein Purification System is based on the remarkable selectivity of patented Ni-NTA (nickel-nitrilotriacetic acid) resin for proteins which contain an affinity tag of six or more histidine residues (consecutive or alternating) — the His tag. This technology allows one-step purification of almost any His-tagged protein from any expression system under native or denaturing conditions. NTA, which has four chelation sites for nickel ions, binds nickel more tightly than metal-chelating purification systems that only have three sites available for interaction with metal ions. The extra chelation site prevents nickel-ion leaching and results in a greater binding capacity and protein preparations with higher purity than those obtained using other metal-chelating purification systems. The QIAexpress system can be used to purify His-tagged proteins from any expression system, including baculovirus, mammalian cells, yeast, and bacteria. ProcedureThe purification of His-tagged proteins consists of 4 stages: cell lysis, binding, washing, and elution (see Protein purification with the Ni-NTA protein purification system). Purification of recombinant proteins using the QIAexpress system does not depend on the 3-dimensional structure of the protein or 6xHis tag. This allows one-step protein purification under either native or denaturing conditions, from dilute solutions and crude lysates. Strong denaturants and detergents can be used for efficient solubilization and purification of receptors, membrane proteins, and proteins that form inclusion bodies. Reagents that allow efficient removal of nonspecifically binding contaminants can be included in wash buffers (see table). Purified proteins are eluted under mild conditions by adding 100–250 mM imidazole as competitor or by a reduction in pH. Reagents compatible with the His/Ni-NTA interaction DenaturantsDetergents Reducing agents Others Salts For long-term storage 6 M Gu·HCl 2% Triton X-100 20 mM β-ME 50% Glycerol 4 M MgCl2 Up to 30% ethanol 8 M Urea 2% Tween 20 10 mM DTT 20% Ethanol 5 mM CaCl2 or 100 mM NaOH 1% CHAPS 20 mM TCEP 20 mM Imidazole 2 M NaCl Applications Ni-NTA matrices provide reliable, one-step purification of His-tagged proteins suitable for any application, including: Structural and functional investigations Crystallization for determination of three-dimensional structure Protein–protein and protein–DNA interaction assays Immunization to produce antibodies Scaling up purification to production scale SpecificationsFeaturesSpecifications ApplicationsProteomics Bead size45–165 µm Binding capacityUp to 50 mg/ml FPLCNo Gravity flow or spin columnGravity flow ProcessingManual/Automated ScaleLarge scale Special featureBatch and column purification Start materialCell lysate Support/matrixSepharose CL-6B Tag6xHis tag Yield100 µg – 100 mgProduct ResourcesYou are not authorized to download the resourceFAQs (16) Sort optionsSort alphabetically What are the compatibilities of different reagents with Ni-NTA matrices?FAQ ID -49ViewHow can I remove imidazole from a protein sample?FAQ ID -91ViewHow can I eliminate contaminating protein in my Ni-NTA 6xHis-tag protein purification?FAQ ID -102ViewShould I use Ni-NTA Agarose in column or batch format for purification of 6xHis-tagged proteins?FAQ ID -147ViewWhat are the features and benefits of the QIAexpress 6xHis Tag System?FAQ ID -193ViewWhat are your recommendations for PCR template preparation for use with the EasyXpress Insect Kit II?FAQ ID -1221ViewHow can I check if any residual proteins remain on the Ni-NTA Agarose matrix after elution?FAQ ID -324View3351 - What is the upper limit for protein size that can bind to Ni-NTA agarose resin?FAQ ID - 3351View3352 - What are the size ranges of Ni-NTA particles?FAQ ID - 3352ViewCan Ni-NTA resins be used to purify protein with an internal His-tag?FAQ ID -496ViewIs it possible to isolate both RNA and recombinant 6xHis-tagged protein from the same sample?FAQ ID -532ViewWhy do you recommend using Triton X for the purification of 6xHis-tagged protein? -100ViewWhat is the difference between Ni-NTA Agarose and Ni-NTA Superflow?FAQ ID -764ViewHow can I be sure that I am harvesting my induced bacterial culture at the best time point for protein expression?FAQ ID -788ViewAre the buffers in the Ni-NTA Fast Start Kit the same as the ones for use with Ni-NTA purchased separately?FAQ ID -791ViewCan I reuse the Ni-NTA Agarose and Ni-NTA Superflow resins?FAQ ID -802ViewShow allShow lessKit Handbooks (2) Sort optionsSort alphabetically The QIAexpressionist - (EN)A handbook for high-level expression and purification of 6xHis-tagged proteinsShow details(EN) - Important Note for Ni-NTA UsersShow detailsShow allShow lessTechnical Information (2) Sort optionsSort alphabetically Critical factors for successful protein crystallization - (EN)Show detailsReliable purification of GST-, His-, and Strep-tagged proteins - (EN)Show detailsShow allShow lessScientific Posters (2) Sort optionsSort alphabetically (EN) - New Ni-NTA Cartridges — the faster way to purer proteinsShow details(EN) - Novel cell-free expression system for synthesis of proteins used in structural analysesShow detailsShow allShow lessQuick-Start Protocols (1) Sort optionsSort alphabetically Ni-NTA Agarose Purification of 6xHis-tagged Proteins from E. coli under Native Conditions (EN)Show detailsShow allShow lessSafety Data Sheets (4) Sort optionsSort alphabetically MSDS Ni-NTA Agarose (25 ml)MSDS Ni-NTA Agarose (100 ml)MSDS Ni-NTA Agarose (500 ml)CofAShow allShow less References 0 fragment fix placeholderCustomers who bought these products also boughtCat No./ID:34964Polypropylene Columns (5 ml)50/pack, 5 ml capacity$189.00Add To CartCat No./ID:34924Polypropylene Columns (1 ml)50/pack, 1 ml capacity$147.00Add To CartCat No./ID:330519 RT² SYBR Green Fluor qPCR Mastermix (25ml) Contains 1 bottle of 25 ml mastermix: for 20 x 96-well RT² PCR Arrays, 12 x 384-well RT² PCR Arrays or 2000 x 25 µl reactionsor 5000 x 10 µl reactions$1,577.00Add To CartCat No./ID:12143QIAGEN Plasmid Midi Kit (25)25 QIAGEN-tip 100, Reagents, Buffers$303.00Add To CartCat No./ID:28704QIAquick Gel Extraction Kit (50)For gel extraction or cleanup of 50 reactions: 50 QIAquick Spin Columns, Buffers, Collection Tubes (2 ml)$124.00Add To CartCat No./ID:27106X4QIAprep Spin Miniprep Kit (1000)For 1000 high-purity plasmid minipreps: 1000 QIAprep 2.0 Spin Columns, Reagents, Buffers, Collection Tubes (2 ml)$1,360.00Add To CartCat No./ID:27104QIAprep Spin Miniprep Kit (50)For 50 high-purity plasmid minipreps: 50 QIAprep 2.0 Spin Columns, Reagents, Buffers, Collection Tubes (2 ml)$96.90Add To CartCat No./ID:28104QIAquick PCR Purification Kit (50)For purification of 50 PCR reactions: 50 QIAquick Spin Columns, Buffers, Collection Tubes (2 ml)$123.00Add To CartCat No./ID:28706X4QIAquick Gel Extraction Kit (1000)For gel extraction or cleanup of 1000 reactions: 1000 QIAquick Spin Columns, Buffers, Collection Tubes (2 ml)$1,879.00Add To CartCat No./ID:28604MinElute Gel Extraction Kit (50)50 MinElute Spin Columns, Buffers, Collection Tubes (2ml)$141.00Add To CartCat No./ID:79254RNase-Free DNase Set (50)1500 Kunitz units RNase-free DNase I, RNase-free Buffer RDD, and RNase-free water for 50 RNA minipreps$113.00Add To CartCat No./ID:12145QIAGEN Plasmid Midi Kit (100)100 QIAGEN-tip 100, Reagents, Buffers$1,133.00Add To CartCat No./ID:76104RNAprotect Tissue Reagent (50ml)50 ml RNAprotect Tissue Reagent for stabilization of RNA in 25 x 200 mg tissue samples$88.60Add To CartCat No./ID:28106QIAquick PCR Purification Kit (250)For purification of 250 PCR reactions: 250 QIAquick Spin Columns, Buffers, Collection Tubes (2 ml)$561.00Add To CartCat No./ID:27106QIAprep Spin Miniprep Kit (250)For 250 high-purity plasmid minipreps: 250 QIAprep 2.0 Spin Columns, Reagents, Buffers, Collection Tubes (2 ml)$422.00Add To CartCat No./ID:77023exoRNeasy Serum/Plasma Starter Kit (20)For purification of total exosome-derived RNA from serum/plasma;includes 20 exoEasy columns (10 Maxi, 10 Midi) and 20 RNeasy MinElute spin columns$650.00Add To CartCat No./ID:12362EndoFree Plasmid Maxi Kit (10)10 QIAGEN-tip 500, Reagents, 10 QIAfilter Maxi Cartridges, Endotoxin-free Buffers$355.00Add To CartCat No./ID:30761Ni-NTA Superflow Cartridges (5 x 5 ml)5 cartridges pre-filled with 5 ml Ni-NTA Superflow: for automated purification of His-tagged proteins using liquid chromatography systems$644.00Add To CartCat No./ID:28004MinElute PCR Purification Kit (50)50 MinElute Spin Columns, Buffers, Collection Tubes (2ml)$141.00Add To CartCat No./ID:28006X4MinElute PCR Purification Kit (1000)1000 MinElute Spin Columns, Buffers, Collection Tubes (2ml)Add To Cart