5-Phospho-D-ribose 1-diphosphate Pentasodium Salt 5-磷酰核糖-1-焦磷酸钠盐

5-Phospho-D-ribose 1-diphosphate Pentasodium Salt 5-磷酰核糖-1-焦磷酸钠盐

货号: JP0260-25MG 品牌: Jinpan 标签: 生化试剂

描述

5-Phospho-D-ribose 1-diphosphate Pentasodium Salt

 5-磷酰核糖-1-焦磷酸钠盐

产品信息

产品名称

产品编号                  规格               价格(元)     

5-Phospho-D-ribose 1-diphosphate Pentasodium Salt        

 5-磷酰核糖-1-焦磷酸钠盐

JP0260-5MG 5mg 540

JP0260-10MG

10mg

990

JP0260-25MG 25mg

1980

JP0260-100MG 100mg

4960

产品描述

5-磷酰核糖-1-焦磷酸(5-Phospho-D-ribose 1-diphosphate, PRPP)是一种天然的中间产物,参与磷酸戊糖途径,引起嘌呤、嘧啶和组氨酸代谢。也是从葡萄糖开始进行植物激素、生物碱和其他二级代谢物的合成途径中的一种中间产物。几种磷酸核糖转移酶(PRTases)利用PRPP为一个底物,将5-焦磷酸基团转移到另一个底物上,正如:通过腺嘌呤PRTase作用于腺嘌呤和PRPP生成腺苷一磷酸(AMP)。

本品是钠盐形式的5-磷酰核糖-1-焦磷酸,CAS NO. 108321-05-7,以冻干粉形式提供,纯度≥75%。

产品特性

  1. CAS NO:108321-05-7
  2. 化学名:5-(dihydrogen phosphate) 1-(trihydrogen diphosphate) α-D-ribofuranose pentasodium salt
  3. 同义名:5-Phospho-D-ribose 1-diphosphate sodium salt; 5-Phosphoribosyl 1-pyrophosphate pentasodium salt; Phosphoribosyl Pyrophosphate pentasodium salt; α-D-Ribosyl diphosphate 5-phosphate pentasodium salt; P-rib-PP; PRPP pentasodium salt; 5-磷酰核糖-1-焦磷酸五钠盐;PRPP五钠盐;
  4. 分子式:C5H8Na5O14P3
  5. 分子量:499.98
  6. 6)  纯度:≥75%
  7. 7)  外观:白色粉末
  8. 8)  溶解性:溶于H2O(50mg/ml)、PBS (pH 7.2)(~10mg/ml)
  9. 化学结构式: 5-Phospho-D-ribose 1-diphosphate Pentasodium Salt 5-磷酰核糖-1-焦磷酸钠盐

保存与运输方法

保存:-20℃干燥保存,2年稳定。

运输:冰袋运输。

注意事项

为了您的安全和健康,请穿实验服并戴一次性手套操作。

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JP0231-25MG Uridine 5′-Diphosphoglucose (UDPG), Disodium Salt Hydrate尿苷5’-二磷酸葡萄糖二钠盐水合物  25mg
JP0258-10MG Guanosine 5′-Diphosphoglucose (GDPG) Sodium Salt鸟苷5’-二磷酸葡萄糖钠盐 10mg
JP0259-5MG Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt尿苷-5’-二磷酸葡糖醛酸三钠盐 5mg
JP0260-5MG 5-Phospho-D-ribose 1-diphosphate Pentasodium Salt 5-磷酰核糖-1-焦磷酸钠盐 5mg

 

 

 

 

规格信息

品牌:

Jinpan

CAS:

108321-05-7

规格:

25mg

货期:

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Coumarin Boronic Acid pinacolate ester (CBE) 香豆素硼酸频那醇酯

Coumarin Boronic Acid pinacolate ester (CBE) 香豆素硼酸频那醇酯 货号: JP4820-100MG 品牌: Jinpan 标签: 细胞生物学研究

描述

Coumarin Boronic Acid pinacolate ester (CBE)

香豆素硼酸频那醇酯

产品关键词

香豆素硼酸频那醇酯(Coumarin Boronic Acid pinacolate ester, CBE);coumarin boronic acid (CBA);Peroxynitrite 过氧亚硝基(ONOO-);Coumarin 6 香豆素6;CAS NO:190788-61-5;

产品信息

产品名称

产品编号     规格    价格(元)

Coumarin Boronic Acid pinacolate ester (CBE) (ONOO-Fluorescent Probe) 香豆素硼酸频那醇酯

JP4820-50MG 50mg 723
JP4820-100MG 100mg

993

JP4820-250MG 250mg

1973

产品描述

香豆素硼酸频那醇酯(Coumarin Boronic Acid pinacolate ester, CBE)是酯化型的香豆素硼酸(Coumarin Boronic Acid, CBA),具更好的溶解性,是一款荧光探针,用来检测过氧亚硝基(ONOO-)、次氯酸(HClO)和过氧化氢(H2O2)[1-4]。过氧亚硝基氧化香豆素硼酸频那醇酯生成荧光产物-7-羟基香豆素(COH),最大激发和发射波长分别是332nm和470nm。

Coumarin Boronic Acid pinacolate ester (CBE)  香豆素硼酸频那醇酯

Fig 1.香豆素硼酸香豆素硼酸频那醇酯(CBE)与过氧亚硝基的反应流程图

产品特征

同义名:CBE; CBA pinacolate ester; Coumarin-7-pinacolboronate;Coumarin 7-Boronic Acid, pinacolate ester; (2-Oxochromen-7-yl)boronic acid;香豆素-7-硼酸频那醇酯;
化学名:7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-1-benzopyran-2-one
CAS NO.:190788-61-5 外观:固体
分子式:C15H17BO4 分子量:272.10
纯度:≥95% 溶解性:溶于DJPO(3mg/ml)、乙醇(1mg/ml)、DMF(10mg/ml),DJPO:PBS (pH 7.2) (1:1) (0.5mg/ml)
化学结构式:Coumarin Boronic Acid pinacolate ester (CBE)  香豆素硼酸频那醇酯

保存与运输方法

保存:-20℃避光干燥保存,至少2年有效。

运输:室温运输。

注意事项

1)  关于化合物溶解性:产品特性内的“≥”表明溶于标示浓度,但饱和溶解度未知。不同批次化合物的溶解度会有差异。

2)  为了让化合物更好的溶解,可通过37℃加热或(和)超声波水浴中震动片刻来处理。若实验所需浓度过大甚至达产品溶解极限,请添加助溶剂助溶或自行调整浓度。

3)  为了您的安全和健康,请穿实验服并戴一次性手套操作。

相关产品

货号 名称 规格
JP4819-50MG Coumarin Boronic Acid (CBA)香豆素硼酸 50mg
JP4820-50MG Coumarin Boronic Acid pinacolate ester (CBE)香豆素硼酸频那醇酯 50mg
JP4821-1G 1,3-diphenylisobenzofuran (DPBF) 1,3-二苯基异苯并呋喃 1g
JP4822-50MG 9,10-Anthracenediyl-bis(methylene)dimalonic Acid (ABDA) 50mg
JP4823-100UG Singlet Oxygen Sensor Green (SOSG)单线态氧荧光探针 100µg
JP4826-25MG [Ru(dpp)3]Cl2(Luminescent oxygen sensor)氧指示探针 25mg

参考文献

[1] Zielonka, J., Sikora, A., Hardy, M., et al. Boronate probes as diagnostic tools for real time monitoring of peroxynitrite and hydroperoxides. Chem. Res. Toxicol. 25(9), 1793-1799 (2012).

[2] Zielonka, J., Zielonka, M., Sikora, A., et al. Global profiling of reactive oxygen and nitrogen species in biological systems. The Journal of Biological Chemisty 287(5), 2984-2995 (2012).

[3] Zielonka, J., Sikora, J., Joseph, J., et al. Peroxynitrite is the major species formed from different flux ratios of co-generated nitric oxide and superoxide. The Journal of Biological Chemisty 285(19), 14210-14216 (2010).

[4] Du, L., Li, M., Zheng, S., et al. Rational design of a fluorescent hydrogen peroxide probe based on the umbelliferone fluorophore. Tetrahedron Lett. 49(19), 3045-3048 (2008).

 

 — —Written/Edited by V. Shallan【版权归金畔生物/JP金畔所有】

规格信息

品牌:

Jinpan

CAS:

190788-61-5

规格:

100mg

货期:

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Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42(HFIP处理)

Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42(HFIP处理) 货号: MP5401-25MG-DZ 品牌: Jinpan 标签: 蛋白研究

描述

Amyloid β Peptide 1-42, Human

人β-淀粉样多肽1-42(HFIP处理)

 

产品信息

产品名称 产品编号 CAS NO 规格         价格(元)
人β-淀粉样多肽1-42(HFIP处理) MP5401-1MG-DZ 107761-42-2 1mg 2480
人β-淀粉样多肽1-42(HFIP处理) MP5401-5MG-DZ 107761-42-2 5mg 4380
人β-淀粉样多肽1-42(HFIP处理) MP5401-25MG-DZ 107761-42-2 25mg 7150

【提示】:点击此处,见Amyloid β Peptide 1-42, Mouse, Rat 大小鼠β-淀粉样多肽 1-42产品信息。

产品描述

Aβ,英文名Amyloid β-protein,中文名β-淀粉样蛋白,具神经营养性和神经毒性,由β-淀粉样前体蛋白(APP)经β-和γ-分泌酶的蛋白水解作用产生的含39-43个氨基酸的多肽,由细胞分泌,沉淀聚积在细胞基质,此过程不仅与神经元的退行性病变有关,而且能激活一系列病理事件,包括星形胶质细胞和小胶质细胞的激活、血脑屏障的破坏和微循环变化等,是导致阿尔茨海默病(AD)患者老年斑周围神经元变性和死亡的主要原因。

Aβ(1-42,1-43)是构成老年斑和神经纤维缠结的主要成分,出现在AD患者的海马体、大脑皮层和杏仁核处。Aβ(39-43)是形成AD和晚期唐氏综合症患者淀粉样蛋白斑的主要组分。Aβ(1-42)可下调bcl-2且提高bax表达水平。此多肽通过作用于p75神经营养素受体来诱导神经死亡。

本品为HFIP处理的人β-淀粉样蛋白1-42,去除了原合成多肽中预先存在的结构(β-折叠、聚集等),此步处理对于控制聚集研究至关重要。

产品特性

1)   CAS NO:107761-42-2

2)  同义名:Amyloid β-Peptide (1-42) (human) (HFIP-treated);β-Amyloid (1-42), human,HFIP-treated; Amyloid β Protein Fragment 1-42,HFIP-treated;

3)   分子式:C203H311N55O60S

4)   分子量:4514.1 g/mol

5)   纯度:≥95%(HPLC)

6)   外观:白色冻干粉末

7)   溶解性:溶于DJPO

8)   单字母序列:DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA

9)   三字母序列:Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-

Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala

保存与运输方法

保存:-20℃避光干燥保存,一年有效。

运输:冰袋运输。

注意事项

1)   本品抗衡离子(Counter Ion)是TFA盐。

2)   本品以冻干粉形式提供,可能因量少不易观察到。请直接加溶剂到瓶子使其溶解。

3)   为了您的安全和健康,请穿实验服并戴一次性手套操作。

相关产品

产品编号 产品名称 规格        
MP5401-1MG Amyloid β Peptide 1-42, Human人β-淀粉样多肽1-42 1mg
MP5401-1MG-DZ Amyloid β Peptide 1-42, Human人β-淀粉样多肽1-42(HFIP处理) 1mg
MP5402-1MG Amyloid β Peptide 1-42, Mouse, Rat大小鼠β-淀粉样多肽1-42 1mg
MP5402-1MG-DZ Amyloid β Peptide 1-42, Mouse, Rat (HFIP-treated)大小鼠β-淀粉样多肽1-42(HFIP处理) 1mg

附录I:不同聚集形态的合成β-淀粉样多肽的制备方法(Preparing Synthetic Aβ in Different Aggregation States)【摘自文献:PMID: 20967580】

Amyloid β Peptide 1-42, Human  人β-淀粉样多肽1-42(HFIP处理)

图:制备寡聚体和纤丝状Aβ1-42的流程汇总图

1)     用HFIP(六氟异丙醇)处理合成多肽,去除现存的聚合物和β-折叠二级结构,形成单体形式的多肽。(Preparation of HFIP-Treated Aβ Peptide Stocks)

Steps 1–7 need to be done in a fume hood.

1.      Prepare a 1 mM Aβ solution by adding HFIP directly to the vial containing lyophilized powder through the rubber septum using a 2.5 mL glass Hamilton syringe with a Teflon plunger and sharp (not blunt-end) needle. For Aβ42, add 2.217 mL to 10 mg peptide (see Note 6).

2.      After the peptide is completely dissolved, pierce the septum with a syringe needle to release the vacuum (see Note 7).

3.      Incubate the Aβ – HFIP solution at room temperature (RT) for at least 30 min (see Note 8).

4.      Decap the glass vial (pliers work well) and remove the rubber septum being careful not to allow the HFIP to come in contact with the septum. Have a rack of 0.5 mL or 1.7 mL micro-centrifuge tubes ready.

5.      Using a positive-displacement repeating pipette, aliquot the solution into 10 μL (0.045 mg for Aβ42) or 100 μL (0.45 mg for Aβ42) aliquots in either 0.5 mL or 1.7 mL microcentrifuge tubes (see Note 9).

6.      Allow HFIP to evaporate in the open tubes overnight in the fume hood.

7.      Transfer tubes to a SpeedVac and dry down for 1 h without heating to remove any remaining traces of HFIP and moisture.

8.      Remove tubes from SpeedVac. The resulting peptide should be a thin clear film at the bottom of the tubes (see Note 10).

9.      Store dried peptide films over desiccant in glass jars at −20°C (see Note 11).

10.    Prior to use, remove peptide film from −20°C freezer and allow sample to come to RT.

11.    Prepare a 5 mM Aβ DJPO stock by adding 20 μL fresh dry DJPO to 0.45 mg Aβ42 peptide (2 μL to 0.045 mg Aβ42). Pipette thoroughly, scraping down the sides of the tube near the bottom to ensure complete resuspension of peptide film (see Note 12).

12.    Vortex well (~30 s) and pulse in a microcentrifuge to collect solution at the bottom of the tube (see Note 13)

2)  未聚集Aβ制备物(Unaggregated Aβ Preparation)

13.    Sonicate 5 mM Aβ DJPO solution for 10 min in a bath sonicator.

14.    Use this preparation as the starting material for unaggregated Aβ (Subheading 3.2), oligomeric Aβ (Subheading 3.3), fibrillar Aβ (Subheading 3.4), “plaque in a dish” (Subheading 3.5), or fluorophore-labeled oligomeric Aβ (Subheading 3.6).

3)寡聚体Aβ制备物(Oligomeric Aβ Preparation)

1.      Start with a tube of freshly resuspended 5 mM Aβ42 in DJPO at RT (see Note 15).

2.      To this Aβ aliquot, add cold phenol-free F-12 cell culture media, diluting to a final concentration of 100 μM Aβ. For example, to 2 μL of 5 mM Aβ in DJPO, add 98 μL cold F-12. Remember to use proper sterile technique. When using F-12 media, avoid prolonged exposure to light and keep F-12 solutions on ice.

3.      Vortex for 15 s, transfer to 4°C and incubate for 24 h.

4.      The expected AFM pattern for this preparation is shown in Fig. 1a, lower left panel.

4)纤丝状Aβ制备物(Fibrillar Aβ Preparation)

1.      Start with a tube of freshly resuspended 5 mM Aβ42 in DJPO at RT (see Note 15).

2.      To this Aβ aliquot, add 10 mM HCl at RT, diluting to a final concentration of 100 μM Aβ. For example, to 2 μL of 5 mM Aβ in DJPO, add 98 μL of 10 mM HCl.

3.      Vortex for 15 s, transfer to 37°C and incubate for 24 h.

4.      The expected AFM pattern for this preparation is shown in Fig. 1a, lower right panel.

5)淀粉样斑制备物(Plaque in a Dish” Preparation)

1.      Start with a tube of freshly resuspended 5mM Aβ42 in DJPO at RT (see Note 15).

2.      To this Aβ aliquot, add 10 mM HCl + 150 mM NaCl, diluting to a final concentration of 100 μM Aβ.

3.      Vortex for 15 s, transfer to 37°C and incubate for 24 h.

4.      The expected AFM pattern for this preparation is shown in Fig. 5a, panel 2

规格信息

品牌:

Jinpan

CAS:

107761-42-2

规格:

25mg

货期:

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Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt 尿苷-5’-二磷酸葡糖醛酸三钠盐

Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt 尿苷-5’-二磷酸葡糖醛酸三钠盐

货号: JP0259-25MG 品牌: Jinpan 标签: 生化试剂

描述

Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt

尿苷-5’-二磷酸葡糖醛酸三钠盐

产品信息

产品名称

产品编号              规格                价格(元)    

Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt    

尿苷-5’-二磷酸葡糖醛酸三钠盐

JP0259-5MG 5mg 540

JP0259-25MG

25mg

1340

JP0259-100MG 100mg

5150

JP0259-250MG 250mg

8500

产品描述

尿苷-5’-二磷酸葡糖醛酸(Uridine 5′-Diphosphoglucuronic Acid, UDPGA)是一种羧酸形成的糖核苷酸,参与许多细胞成分的生物合成。UDPGA是糖基转移酶的糖基供体,用于生物合成多种细胞成分(比如,糖胺聚糖)、一些N-连接和O-连接的聚糖、鞘糖脂和果胶。

本品为三钠盐形式的尿苷-5’-二磷酸葡糖醛酸(UDPGA Trisodium Salt),CAS NO. 63700-19-6,纯度≥98%。

产品特性

化学名:(2S,3S,4S,5R,6R)-6-(((((((2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4- dihydroxytetrahydrofuran-2-yl)methoxy)oxidophosphoryl)oxy)oxidophosphoryl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylate, trisodium salt
同义名:UDP-GlcA trisodium salt, UDPGA trisodium salt, UDPGA -3Na; Uridine-diphosphate-glucuronic acid trisodium salt, Uridine[5′]diphospho[1]-α-D-glucopyranosuronic acid trisodium salt; UDP-GlcA三钠盐;UDPGA三钠盐;尿苷二磷酸葡糖醛酸;尿苷二磷酸-α-葡糖醛酸;5ˊ-二磷酸尿苷葡糖醛酸三钠;尿苷[5ˊ]二磷酸[1]-α-D-葡糖醛酸三钠盐;
CAS NO:63700-19-6 分子式:C15H19N2Na3O18P2
分子量:646.23 g/mol 纯度:≥98%
外观:白色粉末 溶解性:溶于水(50 mg/ml)、PBS (pH 7.2)(~10mg/ml)
化学结构式:Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt 尿苷-5’-二磷酸葡糖醛酸三钠盐

保存与运输方法

保存:-20℃干燥保存,2年稳定。

运输:冰袋运输。

注意事项

为了您的安全和健康,请穿实验服并戴一次性手套操作。

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JP0258-10MG Guanosine 5′-Diphosphoglucose (GDPG) Sodium Salt鸟苷5’-二磷酸葡萄糖钠盐 10mg
JP0259-5MG Uridine 5′-Diphosphoglucuronic Acid (UDPGA) Trisodium Salt尿苷-5’-二磷酸葡糖醛酸三钠盐 5mg

 

 

 

 

规格信息

品牌:

Jinpan

CAS:

63700-19-6

规格:

25mg

货期:

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Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42

Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42 货号: MP5401-1MG 品牌: Jinpan 标签: 蛋白研究

描述

Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42

 

产品信息

产品名称      产品编号           CAS NO.         规格          价格(元)
Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42 MP5401-1MG 107761-42-2 1mg 2050
Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42 MP5401-5MG 107761-42-2 5mg 3650
Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42 MP5401-25MG 107761-42-2 25mg 5950

【提示】:点击此处,见Amyloid β Peptide 1-42, Mouse, Rat大小鼠β-淀粉样多肽1-42产品信息。

产品描述

Aβ,英文名Amyloid β-protein,中文名β-淀粉样蛋白,具神经营养性和神经毒性,由β-淀粉样前体蛋白(APP)经β-和γ-分泌酶的蛋白水解作用产生的含39-43个氨基酸的多肽,由细胞分泌,沉淀聚积在细胞基质,此过程不仅与神经元的退行性病变有关,而且能激活一系列病理事件,包括星形胶质细胞和小胶质细胞的激活、血脑屏障的破坏和微循环变化等,是导致阿尔茨海默病(AD)患者老年斑周围神经元变性和死亡的主要原因。

Aβ(1-42,1-43)是构成老年斑和神经纤维缠结的主要成分,出现在AD患者的海马体、大脑皮层和杏仁核处。Aβ(39-43)是形成AD和晚期唐氏综合症患者淀粉样蛋白斑的主要组分。Aβ(1-42)可下调bcl-2且提高bax表达水平。此多肽通过作用于p75神经营养素受体来诱导神经死亡。

产品特性

  • CAS NO:107761-42-2
  • 同义名:Amyloid β-Peptide (1-42) (human); β-Amyloid (1-42), human; Amyloid β Protein Fragment 1-42;
  • 分子式:C203H311N55O60S
  • 分子量:4514.1g/mol
  • 纯度:≥95%(HPLC)
  • 外观:白色冻干粉
  • 溶解性:溶于1% NH4OH/PBS(1 mg/ml),DJPO(0.4-4 mM)
  • 单字母序列:DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
  • 三字母序列:Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala

保存与运输方法

保存:-20℃避光干燥保存,一年有效。

运输:冰袋运输。

储存液制备

  1. 将低温保存的产品置于室温回温至少20min,低速离心;直接往瓶内加入1% NH4OH溶解粉末(1mg多肽加入70-80μl 1% NH4OH足够);
  2. 之后用1×PBS或者其他合适缓冲液稀释到1mg/ml;【注意】:由于多肽在1% NH4OH溶液不可长期保存,因此必须立即用合适的缓冲液将NH4OH/多肽溶液稀释到约1mg/ml的储存液浓度。
  3. 室温低速漩涡混匀(少于1min)。之后根据单次用量分装冻存,避免反复冻融。
  4. 【注意】:以上储存液制备方法仅做参考,若有特殊要求,可根据经验或参考文献进行其他方法的溶解。

 

注意事项

  1. 本品以冻干粉形式提供,可能因量少不易观察到。请直接加溶剂到瓶子内,漩涡震荡以确保充分溶解;
  2. 为了您的安全和健康,请穿实验服并戴一次性手套操作。

附录I:不同聚集形态的合成β-淀粉样多肽的制备方法(Preparing Synthetic Aβ in Different Aggregation States)【摘自文献:PMID: 20967580】

Amyloid β Peptide 1-42, Human 人β-淀粉样多肽1-42

图:制备寡聚体和纤丝状Aβ1-42的流程汇总图

1)     用HFIP(六氟异丙醇)处理合成多肽,去除现存的聚合物和β-折叠二级结构,形成单体形式的多肽。(Preparation of HFIP-Treated Aβ Peptide Stocks)

Steps 1–7 need to be done in a fume hood.

1.      Prepare a 1 mM Aβ solution by adding HFIP directly to the vial containing lyophilized powder through the rubber septum using a 2.5 mL glass Hamilton syringe with a Teflon plunger and sharp (not blunt-end) needle. For Aβ42, add 2.217 mL to 10 mg peptide (see Note 6).

2.      After the peptide is completely dissolved, pierce the septum with a syringe needle to release the vacuum (see Note 7).

3.      Incubate the Aβ – HFIP solution at room temperature (RT) for at least 30 min (see Note 8).

4.      Decap the glass vial (pliers work well) and remove the rubber septum being careful not to allow the HFIP to come in contact with the septum. Have a rack of 0.5 mL or 1.7 mL micro-centrifuge tubes ready.

5.      Using a positive-displacement repeating pipette, aliquot the solution into 10 μL (0.045 mg for Aβ42) or 100 μL (0.45 mg for Aβ42) aliquots in either 0.5 mL or 1.7 mL microcentrifuge tubes (see Note 9).

6.      Allow HFIP to evaporate in the open tubes overnight in the fume hood.

7.      Transfer tubes to a SpeedVac and dry down for 1 h without heating to remove any remaining traces of HFIP and moisture.

8.      Remove tubes from SpeedVac. The resulting peptide should be a thin clear film at the bottom of the tubes (see Note 10).

9.      Store dried peptide films over desiccant in glass jars at −20°C (see Note 11).

10.    Prior to use, remove peptide film from −20°C freezer and allow sample to come to RT.

11.    Prepare a 5 mM Aβ DJPO stock by adding 20 μL fresh dry DJPO to 0.45 mg Aβ42 peptide (2 μL to 0.045 mg Aβ42). Pipette thoroughly, scraping down the sides of the tube near the bottom to ensure complete resuspension of peptide film (see Note 12).

12.    Vortex well (~30 s) and pulse in a microcentrifuge to collect solution at the bottom of the tube (see Note 13)

2)  未聚集Aβ制备物(Unaggregated Aβ Preparation)

13.    Sonicate 5 mM Aβ DJPO solution for 10 min in a bath sonicator.

14.    Use this preparation as the starting material for unaggregated Aβ (Subheading 3.2), oligomeric Aβ (Subheading 3.3), fibrillar Aβ (Subheading 3.4), “plaque in a dish” (Subheading 3.5), or fluorophore-labeled oligomeric Aβ (Subheading 3.6).

3)寡聚体Aβ制备物(Oligomeric Aβ Preparation)

1.      Start with a tube of freshly resuspended 5 mM Aβ42 in DJPO at RT (see Note 15).

2.      To this Aβ aliquot, add cold phenol-free F-12 cell culture media, diluting to a final concentration of 100 μM Aβ. For example, to 2 μL of 5 mM Aβ in DJPO, add 98 μL cold F-12. Remember to use proper sterile technique. When using F-12 media, avoid prolonged exposure to light and keep F-12 solutions on ice.

3.      Vortex for 15 s, transfer to 4°C and incubate for 24 h.

4.      The expected AFM pattern for this preparation is shown in Fig. 1a, lower left panel.

4)纤丝状Aβ制备物(Fibrillar Aβ Preparation)

1.      Start with a tube of freshly resuspended 5 mM Aβ42 in DJPO at RT (see Note 15).

2.      To this Aβ aliquot, add 10 mM HCl at RT, diluting to a final concentration of 100 μM Aβ. For example, to 2 μL of 5 mM Aβ in DJPO, add 98 μL of 10 mM HCl.

3.      Vortex for 15 s, transfer to 37°C and incubate for 24 h.

4.      The expected AFM pattern for this preparation is shown in Fig. 1a, lower right panel.

5)淀粉样斑制备物(Plaque in a Dish” Preparation)

1.      Start with a tube of freshly resuspended 5mM Aβ42 in DJPO at RT (see Note 15).

2.      To this Aβ aliquot, add 10 mM HCl + 150 mM NaCl, diluting to a final concentration of 100 μM Aβ.

3.      Vortex for 15 s, transfer to 37°C and incubate for 24 h.

4.      The expected AFM pattern for this preparation is shown in Fig. 5a, panel 2

规格信息

品牌:

Jinpan

CAS:

107761-42-2

规格:

1mg

货期:

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