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Ni-IDA Sepharose 6B

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 Catalog No.Package SizePrice
 BD0052-210 mlenquire
 Catalog No.Package SizePrice
BD0052-210 ml$220
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DivBioScience Africa
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Assay Matrix Pty Ltd,
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www.assaymatrix .com
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Gentaur 

Voortstraat 49, 1910 Kampenhout, BELGIUM.
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Sanbio B.V. Frontstraat 2c,5405 PB Uden,The Netherlands 
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FAX: +31(0)413 266 605
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GRUPO IMPEX - IMPEX GROUP
Impex Comércio Internacional Ltda; Bioshop - Materiais para Laboratótios
R.Canada 67/401 - Bairro Sion - Belo Horizonte; M.G. - C.E.P. 30.310-720 Brasil
Tel: (31)3225-2305
Fax: (31) 3227-2422
Email : impexcom@terra.com 
Website:www.impexbrasil.com.br 

BioAspect Inc.
100 College Street, Suite # 322 Toronto ON
M5G 1L5 Canada
Tel: 1-888-609-0366
e-mail: support@bioaspect.com

website: www.bioaspect.com

Nanjing bioworld biotech CO.,Ltd

Room 110, Buliding 6,

Jiangsu Life Science&Technology Innovation Park Weidi road Nanjing, 210046, P, R.China

Tel: +86-25-68037686
E-mail: info@biogot.com

Website:  www.biogot.com

Scintila, s.r.o.
Uvoz 18
586 01 Jihlava, Czech Republic
email: market@scintila.cz
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Nordic BioSite AS
Toldbodgade 18, 5,DK-1253 Copenhagen,Denmark
Tel: +45 4240 8485
Fax: +46 (0)8 756 94 90
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Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
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email       info@divbio.nl 
twitter #divbiobiotech 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
website   www.divbio.nl 
email       info@divbio.nl 
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tebu-bio SRL
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italy@tebu-bio.com

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Netherlands

tebu-bio BV
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netherlands@tebu-bio.com

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tebu-bio 
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UK & Ireland

tebu-bio Ltd
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uk@tebu-bio.com

http://www.tebu-bio.com/cms/741/.html

Nordic BioSite OY
Eteläesplanadi 24,FIN-00130 Helsinki,Finland
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Fax: +46 8 756 94 90
Email : info@biosite.fi 
Website:www.biosite.fi 

 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
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fax  +31 (0)76 201 1229
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AbCys s.a.
5, rue Pierre Chausson, 75010 Paris, France
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FAX: +33 (0)1.44.52.92.69 
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Interchim
211 bis av JF Kennedy - BP1140, 03103 Montlucon cedex - France
tel. 33 (0)4 70 03 88 55
fax : 33 (0)4 70 03 82 60
e-mail : interbiotech@interchim.com  
http://www.interchim.com 

 

tebu-bio
39 Rue de Houdan
78610 Le Perray En Yvelines France
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CliniSciences
183, avenue Georges Clémenceau, 92000 Nanterre - France
Tel: +33 9 77 40 09 09
Fax: +33 9 77 40 10 11
info@clinisciences.com 
www.clinisciences.com 

BIOZOL Diagnostica Vertrieb GmbH 
Obere Hauptstrasse 10b 85386 Eching, German
Tel: +49-89-37 99 666-6
Fax: +49-89-37 99 666-99
Email : info@biozol.de 
Website:www.biozol.de

 
Hölzel Diagnostika Handels GmbH
Hohenzollernring 38
D-50672 Köln
Tel.: +49-(0)221-126 02 66
Fax: +49-(0)221-126 02 67
Email: info@hoelzel.de
Schauen Sie vorbei unter 
http://www.hoelzel-biotech.com  
 
 

TMH Medizinhandel OHG
Medizinischer und Pharmazeutischer Großhandel
Helene-Kropp-Str.1
47166 Duisburg
Tel.: 0203/479721-0
Fax: 0203/479721-88
E-mail: info@tmh-ohg.de 

BioTAG Ltd. 
Tel: 972-52-5522748
Fax: 972-9-8989665

Onwon Trading Limited.
Room 1907, Remington Centre, 23 Hung To Road, Kwun Tong, Hong Kong 
Tel: (852) 2757-7569 
Fax: (852) 2757-7211
Email:info@onwon.com.hk
Website:http://www.onwon.com.hk 

Nordic BioSite AB
Propellervägen 4A,S-183 62 Täby,Sweden
Tel: +46 8 5444 3340
Fax: +46 8 756 94 90
Email : info@biosite.se 
Website:www.biosite.se

Genxbio Health Sciences Pvt. Ltd.
O.No. 6, S-553-554, Ground Floor,
School Block, Shakarpur,
Delhi-110092, India.
Phone no. +91-11-64608275
Fax no. +91-11-43021545
Email: genxbio@gmail.com or sales@genxbio.com
Website: www.genxbio.com 

 

OSCAR DIAGNOSTIC SERVICES PVT. LTD.
307, 3’d Floor, Aggarwal Tower Plot No. 3, L.S.C., CU Block Market Pitam Pura,
New Delhi-110088. INDIA
Phone: + 91-11- 49067453
Fax: +91 8506001695
email: info@odspl.com 
website: www.odspl.com 

 

Clementia Biotech
140, First Floor, RG Mall, Sector - 9 Rohini,
New Delhi - 110085. INDIA
Phone: +91-11-27553034
Email: info@clementiabiotech.com 
Web: www.clementiabiotech.com 

BioTag Ltd.
Palmach 16 st. Kfar Yona
Zip Code 4031016,Israel 

Valter Occhiena S.r.l.
Via Rosta, 7/bis,
10143 Torino, Italy
Tel: +39 011 7716971
Fax: +39 011 7761800
Email: vo@valterocchiena.com  
Website: www.valterocchiena.com   

 

CABRU sas 
via Forlanini, 52,20043 ARCORE (MI).
Tel : +39.039.60.13.988
Fax : +39.039.60.14.284
Email :info@cabru.it 
Website:http://www.cabru.it

 

CliniSciences
Via Maremmana inf. 378, 00012 Guidonia Montecelio - Italy
Telefono : +39 06 94 80 56 71
Fax: +39 06 94 80 00 21
italia@clinisciences.com
www.clinisciences.com 

Funakoshi Co., Ltd. .
9-7 Hongo 2-Chome Bunkyou-Ku, Tokyo 113-0033, Japan 
TEL: +81-3-5684-1620 
FAX : +81-3-5684-1775 
Email:reagent@funakoshi.co.jp
Website:http://www.funakoshi.co.jp


Cosmo Bio Co., LTD.
Toyo-Ekimae Bldg., 2-20, Toyo 2-Chome, Koto-ku, Tokyo 135-0016, Japan
Tel: 81-3-5632-9608 
FAX : 81-3-5632-9965 
Email:kkinno@cosmobio.co.jp 
Website:http://www.cosmobio.co.jp

Arcada for Medical & Laboratory Supplies
AL-Hashimi St, Irbid 21110, Jordan
E-mail: info@arcadajo.com 
Mobile: +962 795 900 474
Telefax: +962 2 7270280

GenomicsOne co., Ltd
DongIn Building 2F, 459 Ogumro, Songpaku, Seoul, Korea (138-813)

Tel:(82)2 430 7722 
Fax:(82)2 430 7724 
Email:info@digenomic.com. 
Website: http://www.digenomic.com. 

 

Kim & Friends Inc.
#1703 Ace High-End Tower 8th, 84, Gasan digital 1-ro,
Geumcheon-gu, Seoul, South Korea ZIP code : 08590
Tel: 82-2-2647-6611
Fax: 82-2-2647-6687
Email: knf_sales@hotmail.com 
Web: www.kimnfriends.co.kr 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
website   www.divbio.nl 
email       info@divbio.nl 
twitter #divbiobiotech 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
website   www.divbio.nl 
email       info@divbio.nl 
twitter #divbiobiotech 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
website   www.divbio.nl 
email       info@divbio.nl 
twitter #divbiobiotech 

 

Sanbio B.V. Frontstraat 2c,5405 PB Uden,The Netherlands 
Tel.: +31(0)413 251 115 
FAX: +31(0)413 266 605
Email : sanbio@sanbio.nl 
Website: sanbio.nl

BIOLUTION RESOURCES (002247091-D)
No. 11, Jalan Bunga Melati 14, Taman Seraya, Ampang, 68000, Selangor, Malaysia
+6016 8988 958
bioservicesmal@gmail.com  
https://biolutionresources.weebly.com 

 

Biomax Scientific Sdn Bhd
12, Jalan Temenggong Taman Desawira
42500 Telok P. Garang Selangor, Malaysia
Tel: 012-519 0930
Fax: 03-3122 7307
Email: siauchi@biomaxsci.com 
Website: www.biomaxsci.com 

 

Bita Lifescience Sdn Bhd.

C-23-05, Dataran 32,No. 2, Jalan 19/1,46300 Petaling Jaya,Selangor Darul Ehsan,Malaysia
Tel: 603-7960-8286
Fax :603-7960-8262
Email:info@bitalifesc.com.my 
Website:www.bitalifesc.com.my 

 

i-DNA Genetics Pte. Ltd.
41 Science Park Road, #01-18/20 Gemini, Singapore Science Park II, SINGAPORE 117610
Tel: (65) 6779 7704
Fax(:65) 6776 0368
Email:thuan@i-genetics.com 
Website:www.i-genetics.com 

Divbio Science Europe
P.O. Box 92,4850 AB Ulvenhout
The Netherlands
phone:+31 (0)76 565 1680
fax: +31 (0)76 201 1229
website: www.divbio.nl
email: frits.stevens@divbio.nl
twitter #divbiobiotech

 

Sanbio B.V. Frontstraat 2c,5405 PB Uden,The Netherlands 

Tel.: +31(0)413 251 115 
FAX: +31(0)413 266 605
Email : sanbio@sanbio.nl 
Website: sanbio.nl 

Nordic BioSite AS
P.b 1681 Vika (Arbins gate 2),N-0253 Oslo,Norway
Tel: 800 10 301
Fax: +46 8 756 94 90
Email : info@biosite.no 
Website:www.biosite.no

 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
website   www.divbio.nl 
email       info@divbio.nl 
twitter #divbiobiotech 

BIOLIM
81-526 Gdynia,Swierkowa 30/6,Poland
Phone: 48 58 746 38 04
Fax: 48 58 746 38 03
Email: info@biolim.pl
Internet: www.biolim.pl 

Genomax Technologies Pte Ltd.
20 Ayer Rajah Crescent, #07-25 S(139964)
Tel: (65) 67771886
Fax:(65) 6777 3090
Email:info@genomax.com.sg 
Website:www.genomax.com.sg

  

Afirmus Biosource Pte Ltd
2 Kallang Avenue #06-32, Singapore 339407
Tel: (65) 6604 6872 / 8163 7412
Fax: (65) 6604 6873
Email: enquiry@afirmus.com
Website: www.afirmus.com 

AntibodyBcn (ABBCN, SL ) 
Parc Científic de Barcelona (PCB). C/Baldiri Reixac 4. ES-08028 Barcelona (SPAIN) 
Tel: 902 220246
Email : info@antibodybcn.com 
Website:www.antibodybcn.com

 

Abyntek Biopharma,S.L.
Parque Tecnológico de Bizkaia, Edificio 504, 48160 Derio - Bizkaia (Spain)
Tel.: (+34) 94 404 8080
Fax: (+34) 94 404 8081
www.abyntek.com 
info@abyntek.com 

Biologist Group d.o.o.
Požeška 67A 11000 Beograd, 11000, Srbija.
Tel: +381 11 3820 094
Fax: +381 62 221 638

Website: www.biologist.rs 

Email: g.mitrovic@biologist.rs

Nordic BioSite AB
Propellervägen 4A,S-183 62 Täby,Sweden
Tel: +46 8 5444 3340
Fax: +46 8 756 94 90
Email : info@biosite.se 
Website:www.biosite.se

 

Divbio Science Europe
P.O. Box 92
4850 AB Ulvenhout The Netherlands
phone  +31 (0)76 565 1680
fax  +31 (0)76 201 1229
website   www.divbio.nl 
email       info@divbio.nl 
twitter #divbiobiotech 

Lucerna-Chem AG 
Abendweg 18,6006 Luzern,Switzerland
Tel: +41 (0)41-420.96.36
Fax: +41 (0)41 420.96.56
Email : lucerna-chem@lucerna-chem.ch 
Website: https://lucerna-chem.ch 

ASIA BIOSCIENCE CO.,LTD.

Contact: Mr. Jimmy Chen

Address: 1 F, No.1, Lane 661, Wenlin Road, Shihlin District, Taipei City 111, Taiwan

Phone: +886-2-8866-3366

Fax: +886-2-2833-1789

http://www.abscience.com.tw

E-mail: info@abscience.com.tw

 

 

BIOTOOLS CO., LTD
TEL:+886-2-26963699
FAX:+886-2-26963689
E-MAIL:info@toolsbiotech.com
http://www.toolsbiotech.com/

 

Li-Tzung Biotechnology, INC
3F.-1,No.211, Hebei 2nd RD., Sanmin District,
Kaohsiung City 807, Taiwan (R.O.C.)
Tel: 886-7-2411512
Fax: 886-7-2411643
E-mail address: li.tzung@msa.hinet.net 

ARP American Research Products, Inc
135 Beaver Street, Suite 204
Waltham, MA 02452
Ph. (800) 832-2611 Fax. (888) 832-2613
www.arp1.com

ALFAGEN BIYOLOJIK URUNLER VE LABORATUVAR MALZEMELERI TIC-SULEYMAN ASLAN 
Kizilay Mah. 483 Sokak No:9 D:2 Bornova/ IZMIR/ TURKEY 
Tel: +90 232 343 27 65
Fax: +90 232 339 89 33 
Email :info@alfagen.com.tr 
Website:http://www.alfagen.com.tr 

2BScientific Ltd
Cherwell Innovation Centre, 77 Heyford Park, Upper Heyford, OX25 5HD, United Kingdom
 Tel: +44(0) 1869 238033
Fax: +44(0) 1869 238034
email: info@2BScientific.com ; sales@2BScientific.com  
Website: www.2BScientific.com  
Link: http://2bscientific.com/~Bioworld 

 

Stratech Scientific Ltd
Cambridge House, St Thomas’ Place, Cambridgeshire Business Park, Ely, CB7 4EX, UK
Phone: +44 (0) 1638 782600
Fax: +44 (0)1638 782606
Email: info@stratech.co.uk 
Web: www.stratech.co.uk 

Bioworld Technology,Inc
1660 South Highway 100, Suite 500
St. Louis Park, MN 55416
Tel: 612-326-3284
Fax: 612-293-3841
Email: info@bioworlde.com
Support email: support@bioworlde.com

Catalog No.BD0052-2
Product NameNi-IDA Sepharose 6B
ApplicationsProtein Related Reagents
Alternative NameRecommended buffers: Native conditions: Binding/wash buffer: 20 mM Tris-HCl, 500 mM NaCl, 5 mM imidazole, pH 8.0. Elution buffer: 20 mM Tris-HCl, 500 mM NaCl, 500 mM imidazole, pH 8.0. Denaturing conditions: Binding/wash buffer: 20 mM Tris-HCl, 8 M urea, 500 mM NaCl, 5 mM imidazole, pH 8.0. Elution buffer: 20 mM Tris-HCl, 8 M urea, 500 mM NaCl, 500 mM imidazole, pH 8.0. Notice: The optimal concentration of imidazole needed in the sample and buffer to obtain the best purity and yield differs from protein to protein. Under native conditions, 20-40 mM imidazole in the binding buffer is suitable for many proteins. 500 mM imidazole in the elution buffer is most often sufficient to completely elute the target protein. Regenerating the medium Note: The medium does not have to be stripped between each purification if the same protein is going to be purified; it is sufficient to recharge the medium after 5–7 purifications, depending on the cell extract volume, target protein, etc. To recharge Ni-IDA Sepharose 6B, first remove residual Ni2+, wash with 5 column volumes 20 mM sodium phosphate, 0.5 M NaCl, 50 mM EDTA, pH 7.4. Remove residual EDTA by washing with at least 5 column volumes of binding buffer followed by 5 column volumes of distilled water before recharging the column. To recharge the water-washed column, load 0.5 column volumes of 0.1 M Ni2SO4 in distilled water. Salts of other metals, chlorides or sulfates, may also be used. Wash with 5 column volumes of distilled water followed by 5 column volumes of binding buffer (to adjust pH) before storage in 20% ethanol.
ClassificationBiogot TM Ni-IDA sepharose 6B has immobilized more Ni2+ than other Ni-IDA ( looks more blue). Our Ni-IDA can couple more protein per ml. Our product is made of GE Sepharose CL gels. The cross-linked form of Sepharose is chemically and physically more resistant than Sepharose itself, offering the same selectivity with better flow characteristics. Cross-linked Sepharose gels are resistant to organic solvents and thus are the choice for separations in organic solvents
FormatLiquid
Contents6% agarose
Storage & Shelf LifeNi-IDA Sepharose 6B is available in 2, 10, 25 and 100 ml lab packs for different requirement. The medium is stored in 20% ethanol at 4°C to 30°C for longer periods of time.
ProcedureSample preparation: The protocol below has been used successfully in our laboratories, but other established procedures may also work. 1. For protein expressed in E. coli or yeast cytoplasm. Dilute the cell paste: Add 5-10 ml of binding buffer for each gram of cell paste. It is essential that the sample and binding buffers contain the same concenctration of imidazole to prevent binding of host cell proteins with exposed histidines.At the same time , remove large particles and high concentration of reagents such as EDTA, amino acids and reducing agents, which can destroy Ni-IDA resins. 2. Enzymatic lysis: 0.2 mg/ml lysozyme, 20 µg/ml DNAse, 1 mM MgCl2, 1 mM PMSF (final concentrations) or other protease inhibitors. The inhibitors must have no effect on the ability of the Ni resin.then Stir for 30 minutes at +4°C. 3. Mechanical lysis: Sonication, homogenization, repeated freeze/thaw or similar techniques. 4. Adjust the pH of the lysate to pH 7.4: Do not use strong bases or acids for pH-adjustment (precipitation risk). 5. Centrifuge the lysate: Transfer to tubes and centrifuge at 12000rpm for 20 minutes at room temperature or +4°C depending on the sensitivity of the protein. 6. Collect supernatants and perform the purification. 7. For proteins secreted into culture medium by yeast, insect, or mammalian expression systems,if have a lot of large volume of supernatant, concentrate the proteins by ammonium sulphate precipitation Dialyze with 1× PBS,then apply to the column, If bit culture supernatant does not contain EDTA, histidine, or any other reducing agents that might affect the Ni column, it can be applied directly to the column. Note: You can also apply unclarified sample to the column (i.e. omitting step 5 above). If so, extend the mechanical lysis somewhat, e.g. sonicate for 10 minutes. Total purification time will increase due to the higher viscosity of the unclarified sample. Purification Procedures: 1. Column preparation. (a) Mix the slurry by gently inverting the bottle several times to completely suspend the resin. (b) Use a pipette to transfer an appropriate volume of Ni resin slurry to the column. Allow the resin to settle and the storage buffer to drain from the column. (c) Equilibrate the column with four bed volumes of binding/wash buffer or until A280 is stable. 2. Purification protein (a) Binding the protein to the resin. Apply the above soluble cleared sample containing His-tagged protein to the column with a flow-rate of 0.5-1 ml per minute. Collect and save the flow-through for analysis. (b) Washing. Wash the column with eight bed volumes of binding/wash buffer or until A280 is stable at the flow-rate of 1 ml per minute. (c) Elution of the target protein. Elute the polyhistidine-tagged protein with five to ten bed volumes of Elution buffer. Collect the elute and dialyze it against 20 mM Tris-HCl pH 8.0 or 1×PBS, pH 7.4, according to the specific application of the target protein. 3. Purification of polyhistidine-tagged proteins (expressed mainly in inclusion bodies) from E. coli under Denaturing conditions: (a) Resuspend the cell pellet in 1× PBS (about 5 ml per ml of pellet), and disrupt cells by sonication as described above. (b) Collect inclusion bodies by centrifuging the lysate at 12,000 rpm for 10 minutes. Wash inclusion bodies with 1× PBS several times if necessary. (c) Solubilize the inclusion bodies in binding/wash buffer (about 5 ml per ml of pellet), and incubate for 30-60 minutes at room temperature. Homogenization or sonication may be necessary to fully solubilize the pellet. (d) Centrifuge at 12,000 rpm for 30 minutes to remove any remaining insoluble material. Carefully transfer supernatant to a clean tube without disturbing the pellet and load it on the Ni column pre-equilibrated with binding/wash buffer. (e) Wash the column with binding/wash buffer for 3-4 times until the absorption at 280 nm is close to zero. (f) Elute with minimal volume of elution buffer. Note: The process recommended here is the purification of protein from inclusion body, the eluted protein from this process may need to be refolded to obtain the active and soluble protein.
Research useFor research use only, not for use in diagnostic procedures.
Expiration DateThe validity of reagents in kit is six months (Please use within the validity period ).

Background

Purifying histidine-tagged recombinant proteins by immobilized metal affinity chromatography (IMAC) continues to grow in popularity. Nickel (Ni2+) is the most commonly used metal ion in IMAC purifications. Ni-IDA Sepharose 6B consists of 90 μm beads of highly cross-linked agarose, to which a chelating ligand has been immobilized. This chelating group has then been charged with nickel (Ni2+) ions. To compare with other Nickel affinity chromatography, Ni-IDA has higher protein-binding capacity. It has low Ni2+ leakage and is compatible with a wide range of additives used in protein purification. Its high flow properties make it excellent for scale-up and gravity flow column purification.

References for Ni-IDA Sepharose 6B

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