高品質蛋白純化樹脂  | PurKine™ His-Tag Ni-NTA Column

PurKine™ His-Tag 蛋白純化試劑盒 (Ni-NTA) | PurKine™ His-Tag Protein Purification Kit (Ni-NTA)貨號KTP2001

 50 mg  6×His-tagged protein/mL  平均珠粒尺寸為45-165 µm   1ml/1mlx5

• PurKine™ His-Tag Purification Nickel Column
• Phosphate-Buffered Saline (PBS,10X)
• Imidazole (2M)

PurKine™ His-Tag 純化系統(PurKine™ His-Tag Purification system)基於創新的高容量 IMAC 基質(IMAC matrix)，可方便地從總裂解物(total lysates)中一步純化 His-tag 蛋白。專有的離子螯合化學(ion-chelate chemistry)確保與常用的還原劑（reducing agents, 如 DTT）、螯合金屬蛋白酶抑制劑（chelating metalloprotease inhibitors, 如 EDTA）以及各種緩衝物質(buffer substances)和鹽條件(salt conditions)具有非凡的相容性。該產品組合提供多種螯合基團（IDA、NTA 或 Super NTA）、交聯瓊脂糖（交聯 4% 或 6% 瓊脂糖）、離子（Nick、Colbat、銅或非帶電離子）和兼容成分，以實現優化最大蛋白質產量、穩定性和溶解度的過程。測試還證實，在至少五次重複使用後性能沒有下降。 PurKine™ His-Tag 純化樹脂也可作為預裝離心柱(prepacked spin column)和試劑盒(kit)形式提供。

PurKine™ His-Tag 蛋白純化試劑盒(PurKine™ His-Tag Protein Purification Kit, KTP2001)可在各種規模的重力柱(gravity column)程序中有效純化高水平過表達的 His-tag 融合蛋白。試劑盒中的樹脂(Resin) 由 90 μm 高度交聯的 4% 瓊脂糖珠(highly cross-linked 4% agarose)組成，與硝酸三乙酸 (Nitilotriacetic acid , NTA) 偶聯。然後螯合基團帶有鎳離子 (nickel ions , Ni2+)。測試證實其性能等於或超過其他供應商的流行 Ni-NTA 樹脂(Ni-NTA resins)，並且在至少五次重複使用後性能沒有下降。

Product name	PurKine™ His-Tag Protein Purification Kit (Ni-NTA)
Applications notes	PurKine™ His-Tag Protein Purification Kit effectively purifies high levels of overexpressed His-tagged fusion proteins in gravity column procedures at a variety of scales. The Resin in kit consists of 90μm beads of highly cross-linked 4% agarose, to which Nitilotriacetic acid (NTA) has been coupled. The chelating group has then been charged with nickel ions (Ni2+). Tests confirm that performance equals or exceeds popular Ni-NTA resins from other suppliers, and no decrease in performance occurs after at least five repeated uses.
Alternative	Ni-NTA; His resin; Ni-NTA resin; His tag resin
Kit components	• PurKine™ His-Tag Purification Nickel Column • Phosphate-Buffered Saline (PBS,10X) • Imidazole (2M)
Features & Benefits	• High capacity—More than 50 mg of 6xHis-tagged protein per milliliter of resin. •Versatile—For purification under native or denaturing conditions. • Cost-effective—No performance decrease after at least five repeated uses of the same batch of resin. • Flexible—available in multiple formats including bulk resin, spin columns and complete kits. • Easy to use— Pre-formulated buffers available for kit formats.
Storage instructions	Stable for one year at 2-8°C from date of shipment. Do not freeze.
Shipping	Blue ice
kground	PurKine™ His-Tag Purification system is based on innovative high-capacity IMAC matrix for convenient single-step purification of His-tag proteins from total lysates. Our propriety ion-chelate chemistry ensures extraordinary compatibility with commonly used reducing agents such as DTT, chelating metalloprotease inhibitors such as EDTA, and a wide range of buffer substances and salt conditions. The portfolio provides wide choice of chelating group (IDA, NTA or Super NTA), cross-linked agarose (crosslinked 4% or 6% agarose), ions (Nick, Colbat, Copper, or non ion charged) and compatible ingredients to allow optimization of process for maximum protein yield, stability and solubility. Tests also confirm that no decrease in performance occurs after at least five repeated uses. The PurKine™ His-Tag Purification resin is also available as prepacked spin column and kit formats.
Alternative	Ni-NTA; His resin; Ni-NTA resin; His tag resin

Sample Preparation

Ⅰ Protein extraction from bacteria or yeast

1. Transfer bacteria or yeast culture medium to centrifugal tube after induced protein expression, centrifuge 15 min at 7,000 rpm to collect bacteria or yeast. 2. Mix with 10 times volume of Lysis buffer (collected bacteria or yeast:Lysis buffer=1:10), add PMSF with final concentration 1mM. Lysozyme is recommended to be added with a final concentration of 0.2-0.4 mg/mL. 3. Resuspend the cells. Add 10 μg/ml RNase A and 5μg/ml DNase I in case the cells’ concentration is high. Mix and sonicate the suspension in ice, keep the lystes to be clean. 4. Transfer the clean lysate to a new centrifuge tube, centrifuge for 20-30 min at 10,000 rpm, 4℃. Take the supernatant, store on ice or -20℃.

Ⅱ Soluble protein extraction from yeast, insect (昆蟲) or mammalian cells

1. Transfer cell culture medium to centrifugal tube, centrifuge 10 min at 5,000 rpm to collect suspension. Suspension should be dialyzed with Lysis buffer in case there’re EDTA, histidine and other reductants in suspension.

2. Large volume of suspension should be concentrated by Ammonium sulfate fractional precipitation method, following be dialyzed with Lysis buffer.

Ⅲ Inclusion-body protein extraction for purification in denaturing conditions

1. Transfer culture medium to centrifugal tube, centrifuge 15 min at 7,000 rpm to collect precipitation. Remove the suspension.

2. Mix with 10 times volume of Lysis buffer, such as collected precipitation: Lysis buffer= 1:10 (W/V). Lysis buffer does not contain 8 M Urea. Resuspend the precipitation. Mix and sonicate the suspension in ice.

3. Transfer the lysate to a new centrifuge tube, centrifuge for 20-30 min at 10,000 rpm, 4℃. Dispose the supernatant, Repeat step 2 and 3 once again.

4. Mix with 10 times volume of Lysis buffer, such as collected precipitation: Lysis buffer=1:10 (W/V). Resuspend the lysate for purification in denaturing conditions.

Reagent Preparation

It is recommended to filter all buffers before use by passing through a 0.22 μm or 0.45 μm filter. For most proteins, the following buffer are recommended:

Note: Sometimes overexpressed proteins are sequestered in inclusion bodies. Inclusion bodies of His-tagged proteins can be solubilized in 8M urea or 6M guanidine. We recommend you use the buffer below (Reagents below are not supplied in the kit)： For imidazole elution method prepare the following buffers: Lysis buffer: 8 M Urea, 50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, pH 8.0; Wash buffer: 8 M Urea, 50 mM NaH2PO4, 300 mM NaCl, 20 mM imidazole, pH 8.0; Elution buffer: 8 M Urea, 50 mM NaH2PO4, 300 mM NaCl, 250 mM imidazole, pH 8.0. For pH elution method prepare the following buffers: Lysis buffer: 8 M Urea, 100 mM NaH2PO4, 100 mM Tris·HCl, pH 8.0; Wash buffer: 8 M Urea, 100 mM NaH2PO4, 100 mM Tris·HCl, pH 6.3; Elution buffer: 8 M Urea, 100 mM NaH2PO4, 100 mM Tris·HCl, pH 4.5.

Procedure for Sample Purification

1. Fix Column. Move the top and bottom stopper, and let the storage buffer drain away.

2. Add 5 resin-bed volume Lysis Buffer to the column. Equilibrate the column (make the Ni-NTA Resin in the same buffer system as the target protein to protect the protein). Allow buffer to drain from the column.

3. Add the prepared protein extract to the resin. (In order to improve the recovery rate of the target protein, the adding speed was controlled to ensure the full contact between the target protein and Ni2+). Note: Collect the flow-through which can be analyzed by SDS-PAGE. When problems arise, it is easier to find solutions.

4. Add 10-15 resin-bed volume Wash Buffer to the column to remove the non-specific adsorption protein. Pay attention to collecting the flow-through.

5. Add 5-10 resin-bed volume Elution Buffer to the column to wash the target protein. The collected eluate is the target protein solution.

6. Add 3 resin-bed volume Lysis Buffer and 5 resin-bed volume deionized water to the column in turn to equilibrate the Ni-NTA Resin. Store resin in an equal volume of PBS containing 20% ethanol at 4-30℃ to prevent the resin from being contaminated by bacteria.

7. The flow-through, eluted protein and prepared protein extract can be directly analyzed by SDS-PAGE to test the purification effect.

Cleaning-in-Place (CIP)

When the back pressure is too high or obvious contamination appears on the resin during the use of the resin, it needs to be cleaned-in-place (CIP). It is recommended to follow the steps below to remove residual contaminants on the resin, such as precipitated proteins, hydrophobins, and lipoproteins.

To remove strongly bound hydrophobic proteins, lipoproteins and lipids: Wash the column using 5-10 resin-bed volumes of 30% isopropanol contacting for 15-20 min. Or apply 2 resin-bed volumes of acidic or alkaline solution containing detergent (i.e. 0.1 M acetic acid solution contains 0.1-0.5% non-ionic detergent), for 1-2 hours. Finally wash the column with 10 resin-bed volumes of distilled water.

To remove the proteins engaged with ionic interaction: Wash the column with 1.5 M NaCl for 10-15 min. Finally wash the column with 10 resin-bed volumes of distilled water.

Ni-NTA Resin Regeneration

In general, The Ni-NTA resin may be used at least five times before it becomes necessary to recharge them with metal ions. When the back pressure is too high or the capacity significantly lower, it needs to strip the metal ions and recharge the resin as the following procedure:

1. Wash resin with 5 resin-bed volumes of deionized water;

2. 100 mM EDTA (pH 8.0), 5 resin-bed volumes;

3. Wash resin with 10 resin-bed volumes of deionized water;

4. Wash resin with 5 resin-bed volumes of 0.5 M NaOH and stay for 10-15 min;

5. Wash resin with 10 resin-bed volumes of deionized water;

6. 100mM NiSO4, 3-5 resin-bed volumes;

7. Wash resin with 10 resin-bed volumes of deionized water; After regeneration, the medium can be used immediately or store in PBS containing 20% ethanol at 4℃.

活細胞和死細胞雙染色套組  |  CCK-8細胞增殖和細胞毒性試劑盒  | LDH細胞毒性測定試劑盒 | 細胞衰老檢測試劑盒（β-半乳糖苷酶 | 細胞增殖EdU Image試劑盒 

BRAND/AGENCY/台灣代理商/ Abbkine台灣代理商

https://www.abbkine.com/

Abbkine Scientific Co., Ltd由一群志同道合的科學家和行銷專家，在2012年成立於美國加利福尼亞州，隨著亞洲市場的迅猛增長，Abbkine將總部搬遷至中國。結合了Abbkine美國技術和中國製造兩方面的巨大優勢，Abbkine致力於提供創新的、高品質的、買得起的實驗試劑盒、蛋白質、抗體以及其它研究工具，以期成為生命科學研究發展，藥物研發等領域的關鍵推動者。

發展歷程

Abbkine致力於提供創新的、高品質的、買得起的實驗試劑盒、蛋白質、抗體以及其它研究工具以期成為生命科學研究發展，藥物研發等領域的關鍵推動者。Abbkine以特色的蛋白質生產與檢測試劑起家，致力於創新和研發各類抗體、生化試劑、蛋白質和實驗試劑盒，以期望為全球科研工作者提供革新的解決方案。我們很自豪地為客戶提供如下特色產品線，並會一如既往地開發新型的、高品質的、買得起的生命科學研究和診斷試劑產品。2017年檢測細胞因數和激素的ELISA試劑盒應運而生: 根據市場需求，開發EliKine™系列ELISA試劑盒，涵蓋各類細胞因數和激素熱門指標，如白介素（IL）、腫瘤壞死因數（TNF）、干擾素（IFN）等明星級偶聯標記試劑盒全球發售。2018年憑藉先進的蛋白質偶聯技術和嚴格的品質控制，我們提供LinKine™偶聯標記試劑盒，在抗體、蛋白質、多肽或者其它生物分子上可直接標記HRP，Biotin，FITC，Cy3，AbFluor™以滿足科研應用，藥物研發以及診斷試劑盒開發的需求。2019年生化、細胞類產品線逐步擴充搭建細胞產品研發平臺，細胞狀態檢測、細胞染色、細胞提取、生化試劑盒產品得到越來越多全球客戶的認可。

活細胞和死細胞雙染色套組  |  CCK-8細胞增殖和細胞毒性試劑盒  | LDH細胞毒性測定試劑盒 | 細胞衰老檢測試劑盒（β-半乳糖苷酶 | 細胞增殖EdU Image試劑盒 

細胞增殖&毒理&衰老 (8)

產品貨號	產品品項	反應	測量儀器
貨號KTA1001	活&死細胞雙染色試劑盒	100個反應	活細胞綠色螢光染劑Ex/Em = 488/530 nm; 死細胞紅色螢光染料(Ex/Em = 535/617)
貨號KTA3030	細胞衰老檢測（β-半乳糖苷酶）	100個反應	光學顯微鏡
貨號KTA1020	CCK-8細胞增殖和細胞毒性試劑盒	1000個反應	吸收光450nm
貨號KTA2030	EdU Image細胞增殖試劑盒 (BrdU替代法)	100個反應	AbFluor 488 azide (Ex/Em = 501/525 nm)
貨號KTB1110	（LDH）乳酸脫氫酶測定試劑盒	96個反應/	λmax= 450 nm 1- 20 U/mL of LDH
貨號 KTA2020	細胞週期染色檢測	50個反應/100個反應	Ex/Em=535/615 nm.
貨號KTA1030	LDH細胞毒性測定試劑盒	48個反應/ 96個反應	OD 565 nm 10,00-100,0000 cells per well
貨號BMD00064-50UG	Calcein AM鈣黃綠素活細胞螢光染劑	50ug	λEX/λEm: 494/517 nm (pH 8) (calcein)

 細胞凋亡&細胞染色 (8)

產品貨號	產品品項	反應	測量儀器
貨號 KTA2010	TUNEL細胞凋亡檢測試劑盒	50個反應	螢光酶標儀、螢光顯微鏡或流式細胞儀。FITC通道（Ex / Em = 490nm / 520 nm）
貨號KTA0002	Annexin V-AbFluor™488細胞凋亡檢測試劑盒	50個反應	Annexin V- AbFluor™ 488: Abs/Em = 491/517 nm; PI: Abs/Em = 535/617 nm (with DNA)
貨號KTA4001	JC-1線粒體膜電位測定試劑盒	20個反應	（聚結的JC-1，Ex / Em = 585/590 nm）發出綠色螢光（JC-1單體，Ex / Em = 510/527 nm） JC-1 Stain CCCP (10 mM)  Assay Buffer (5×)
貨號BMD00063	DAPI Staining Solution	10MG	λEx/λEm: 358/461 nm (with DNA)
KTD102-EN	凋亡細胞分析(Annexin V法和TUNEL法)	40個分析/20個分析
貨號KTC4100	Pro活細胞微管蛋白染色	50個反應/250個反應	Ex/Em = 500/520 nm
貨號KTC4001	Abbkine 細胞膜染色	100個反應/500個反應	Ex/Em = 484/501 nm
貨號KTC4003	TraKine™粒線體染色	100個反應	Ex / Em = 490/523 nm增殖(proliferating)和非增殖細胞(non-proliferating cells)

DNA Damege損傷/細胞Stress (8)

產品貨號	產品品項	反應	測量儀器
貨號KTB1050	脂質過氧化檢測試劑盒(MDA)	96個反應	OD 532 nm 1-100 µM/L
貨號KTB1910	活性氧（ROS）檢測分析試劑盒	50個反應/100個反應	DCFH-DA (10 mM)
貨號KTB1400	一氧化Dan（NO）檢測試劑盒	48個反應/ 96個反應	colorimetric (540 nm) 檢測範圍：1-100 µM
貨號KTB1080	超氧陰離子清除測定試劑盒	48個反應/ 96個反應	Superoxide anion scavenging rate D% =（∆∆ODControl – ∆∆ODSample）÷ ∆∆ODControl ×100%
貨號 KTB1091	羥基自由基清除能力比色測定試劑盒	48個反應/ 96個反應	吸收光 520nm
貨號KTB1041	CheKine™ 過氧化氫 (H2O2) 檢測試劑盒	96個反應/ 480個反應	OD 580 nm : 1 µM-100 µM
貨號KTB1210	CheKine™ 超氧陰離子比色測定試劑盒	48個反應/ 96個反應	absorption peak at 540nm 0.0125-0.5 mg/mL
貨號KTB1020	NAD/NADH 分析試劑盒	48個反應	OD565 nm 96 孔板測定中的檢測限值為 0.78 μM 線性最高 50 μM NAD+/NADH。

抗氧化分析試劑 (8)

產品貨號	產品品項	反應	測量儀器
貨號KTB1030	超氧化物歧化酶（SOD）活性測定試劑盒	48個反應 96個反應	200-3.13 U/ml
貨號KTB1040	Catalase過氧化氫酶活性檢測試劑盒	96個反應	OD 540 nm 2-75 µM
貨號KTB1500	總抗氧化劑能力TCA分析	96個反應/ 480個反應	593 nm 偵測範圍: 0.15-3 mM
貨號 KTB1660	硫氧還蛋白過氧化物酶（TPX）檢測	48個反應/ 96個反應	240nm
貨號KTB1600	穀胱甘肽(Glutathione)檢測	48個反應 96個反應	412 nm  2-200 µg/mL
貨號KTB3010	CheKine™ 花青素還原酶（ANR）活性比色測定試劑盒	96個反應	absorption at 340 nm
貨號KTB1530	黃酮類化合物檢測套組	48個反應 96個反應	absorption peak at 502 nm 0.156-10 mg/g
貨號KTB1140	多酚氧化酶檢測套組	48個反應 96個反應	absorption peak at 525 nm

膽固醇代謝/代謝試驗 (8)

產品貨號	產品品項	反應	測量儀器
貨號KTB1510	CheKine™尿酸（UA）比色分析試劑盒	48個反應/ 96個反應	吸收光505 nm Standard: 5 μmol/mL.
貨號KTB1300	Glucose葡萄糖測定試劑盒	96個反應	吸光值630nm 4- 300 mg/dL)
貨號KTB1340	CheKine™ 肝醣/糖原測定試劑盒	96個反應
貨號KTB2220	TC總膽固醇檢測試劑盒	48個反應/ 96個反應	500nm Standard: 5 μmol/mL
貨號KTB2250	CheKine™ 微量高密度脂蛋白膽固醇 (HDL-C) 檢測試劑盒	48個反應/ 96個反應	Absorption peak at 500nm Detection range: 0.078-5 mmol/L(The Detection range corresponds to the standard, and the actual content of sample is 0.156-10 mmol/L）
貨號KTB2260	CheKine™ 微量低密度脂蛋白膽固醇 (LDL-C) 檢測試劑盒	48個反應/ 96個反應	Absorption peak at 500nm Detection range: 0.078-5 mmol/L(The Detection range corresponds to the standard, and the actual content of sample is 0.117-7.5 mmol/L）
貨號KTB1390	CheKine™ 澱粉分支酶 (SBE) 活性比色測定試劑盒	48個反應/	absorption at 660 nm
貨號KTB1070	CheKine™黃嘌呤氧化酶檢測	96個反應	OD 450 nm 線性範圍15.6 -500 mU/

生化檢測試劑盒 (8)

產品貨號	產品品項	反應	測量儀器
貨號KTB1700	組織和血液鹼性磷酸酶（ Alkaline Phosphatase, AKP / ALP）比色測定	96個反應/	吸收光510 nm
貨號KTB1542	Tannase(TAN) 活性比色測定試劑盒	48個反應/	absorbance at 270 nm 0.3125-20 μmoL/mL
貨號KTB1541	CheKine™ 單寧比色測定試劑盒 | CheKine™ Tannin Colorimetric Assay Kit	48個反應/	absorbance at 760 nm 0.0156-1 mg/mL
貨號KTB1290	CheKine™ 烏頭酸酶 (ACO) 活性比色測定試劑盒	48個反應/	absorption at 340 nm.
貨號KTB1560	醇醯基轉移酶（AAT）活性比色檢測試劑盒	48個反應/ 96個反應	absorption at 412 nm.
貨號KTB1070	CheKine™黃嘌呤氧化酶檢測	96個反應	OD 450 nm 線性範圍15.6 -500 mU/
貨號KTB1380	β-澱粉酶活性比色測定	48個反應	absorption peak at 540 nm 0.0156-1 mg/mL
貨號KTB1310	葡萄糖氧化酶活性測定	48個反應	OD 580 下測量 偵測範圍: 0.05 – 2 U/L

發炎反應ELISA分析

產品貨號	產品品項	反應	測量儀器
貨號KET7013	IL-6 ELISA Kit試劑盒	96個反應/	450 nm 偵測範圍: 250pg/mL-16000 pg/mL, 最低靈敏度100 pg/mL
貨號KET6032	TNF–α Alpha型腫瘤壞死因子 ELISA試劑盒	96個反應/	450 nm 7.8 pg/ml-500 pg/ml
貨號KET6033	VEGF血管內皮生長因子ELISA試劑盒	96個反應/	450 nm 偵測範圍: 31.25 pg/mL-2000 pg/mL
KET7013	IL-22 ELISA試劑盒	96個反應/
貨號KET7003	IFN-γ（干擾素-γ）ELISA試劑盒	96個反應/	31.25 pg/ml- 2000 pg/ml
貨號KET6019	IL-10 ELISA試劑盒	96個反應/	2.35 pg/ml-150 pg/ml
貨號KET7002	GM-CSF ELISA試劑盒	96個反應/	7.8 pg/mL-500 pg/mL
貨號KET7005	白介素-1βELISA試劑盒	96個反應/ 96 X5盤	Microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm. 偵測範圍: 15.6 pg/ml-1000 pg/ml

重組蛋白&單多株抗體

產品貨號	產品品項	反應	測量儀器
貨號PRP1012	Human IL-6 protein重組蛋白	5ug	human IL-6 (NP_000591.1) (Val 30-Met 212)/ E. coli
貨號PRP100273	Human IL-4 protein重組蛋白	20ug	ED50 for this effect is 0.05-0.2 ng/mL. The specific activity of Recombinant Human IL-4 is approximately 3.2 × 10E4 IU/μg.
貨號PRP100159	Human EGF protein重組蛋白	100ug
貨號PRP1012	Human IL-6 protein重組蛋白	5ug	human IL-6 (NP_000591.1) (Val 30-Met 212)/ E. coli
貨號ABP52783	GAPDH Polyclonal Antibody	30ul	Human, Mouse, Rat。可應用於ELISA, IHC-P, WB實驗。
貨號A21010	HRP, Goat Anti-Mouse IgG	100ul
貨號A21020	HRP, Goat Anti-Rabbit IgG	100ul
貨號A23220	Dylight 488, Goat Anti-Rabbit IgG	100ul

抗體蛋白純化&其它