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Ni TED Beads 6FF
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Ni TED Beads 6FF

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1st 年

連絡先担当者 Ying

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タイトルまたは説明

Ni TED Beads 6FF CAT:N30250 Price: $62 5ml; $137-10ml (negotiable)
Storage:2-8℃
 
Description
LABLEAD Ni affinity chromatography medium belongs to a kind of metal rock medium, by the high velocity of agarose for matrix, with IDA, NTA, TED as ligands, and rock and metal ions, Ni2 +, and formed a kind of affinity chromatography medium for integration in a different way, Ni2 + ion binding force there is also a slight difference, It shows different selectivity in the purification process of different proteins and is widely used for the isolation and purification of His-tag proteins. Among them, Ni-TED beads 6FF can tolerate 100mM EDTA and 10mM DTT. The target protein was purified directly in the presence of EDTA and DTT. The cleaning and regeneration of this medium was simple, and the NaOH cleaning was performed directly without nickel removal.
Technical index:
NAME Ni-TED beads 6FF
LIGAND Three (methyl) ethylenediamine (TED)
MATRIX 6% highly crosslinked agarose
Amount of chelation 30~60umol/mL
CAPACITY(PER ml) 30mg His-tag protein
mean grain size 90um,distribute in45~165um
Recommended flow rate 150~15000px/h (Select the appropriate flow rate according to the column specifications)
Maximum pressure resistance 0.3MPa
pH stability 3~12(work) 2~14(washing)
chemical stability 0.01M hydrochloric acid, 0.01M sodium hydroxide (one week);
100mMEDTA, 10mM DTT, 1M sodium hydroxide, 8M urea, 6M hydrochloric acid (24 hours);
100mMEDTA, 0.5mmic (2 hours); 30% isodiol (20 minutes).
 
 
Instructions for operation
The Ni-TED Chromrose 6FF can be packed into a medium pressure chromatographic column in the laboratory to scale up production. The filler was packed into the chromatographic column, and the appropriate chromatographic column and column height were selected according to the sample volume and filler loading.
Buffer preparation
The buffer used should be prepared with high purity water, and it is recommended to filter with 0.45um filter membrane before use.
Equilibration buffer: 0.2M NaCl, 50mM Tris-Hcl, 0.1mM EDTA, PH=7.4
Rinse buffer: 0.2 M NaCl, Tris Hcl - 50 mM, 0.1 mM EDTA, 30 mM imidazole, PH = 7.4
Elution buffer: 0.2M NaCl, 50mM Tris-Hcl, 0.1mM EDTA, 500mM imidazole, PH=7.4
 
1. Sample preparation
The solution of the sample should be consistent with the equilibrium solution, which can be used for lysis, dialysis/ultrafiltration /G25 buffer replacement.
Samples were filtered (mean particle size < 45um, filtered with 0.22um; 45um< average particle size < 165um, filtered with 0.45um; Mean particle size >165um, filtered with 0.8um).
2. The purified sample
3.1 Rinse out the storage buffer with 3~5CV pure water;
3.2 with the balance of 5 ~ 10 CV buffer balance chromatography column, to outflow liquid conductivity and pH unchanged (consistent with fluid balance);
3.3 Sample loading by pump or sample loading ring;
3.4 After loading, the samples were rinsed with elution buffer for 10 to 15CV until the baseline was stable
3.5 One-step or linear gradient elution was performed with elution buffer. Step elution generally 55 CV, gradient elution can use a small gradient, such as 20 times column volume to separate different bonding strength of protein;
3.6 After washing the chromatography column successively with 3CV equilibration buffer, 5CV pure water, and 2CV 20% ethanol, the column was stored at 2-8 °C.
3. In the cleaning
When the backpressure of the filler is found to be too high or obvious pollution occurs on the filler during use, it needs to be cleaned in place.
3.1 remove the combination of strong hydrophobic protein, lipoprotein and lipids
Method 1: use 30% isopropanol to clean 5~10CV, contact time of 15~20min can remove such pollutants, and then use 10CV pure water to clean;
Method 2: use the 0.3 M or 0.5 M NaOH solution washing packing 3 CV, and then clean with 1015 CV of pure water.
4.2 Removal of ion-bound proteins
Using 1.5 M NaCl solution cleaning 10 ~ 15 min, reoccupy 10 CV pure water cleaning, cleaning use 2 CV of 20% ethanol wash good pillar at 2~8°C to save
image
 

国: China
モデル番号: -
离岸价格: Get Latest Price
ロケーション: China
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製品グループ : Protein Biology
Ni TED Beads 6FF

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