Amyloid β Peptide 1-42, Human

人β-淀粉樣多肽1-42（HFIP處理）

產(chǎn)品信息

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產(chǎn)品描述

Aβ，英文名Amyloid β-protein，中文名β-淀粉樣蛋白，具神經(jīng)營(yíng)養性和神經(jīng)毒性，由β-淀粉樣前體蛋白（APP）經(jīng)β-和γ-分泌酶的蛋白水解作用產(chǎn)生的含39-43個(gè)氨基酸的多肽，由細胞分泌，沉淀聚積在細胞基質(zhì)，此過(guò)程不僅與神經(jīng)元的退行性病變有關(guān)，而且能激活一系列病理事件，包括星形膠質(zhì)細胞和小膠質(zhì)細胞的激活、血腦屏障的破壞和微循環(huán)變化等，是導致阿爾茨海默?。ˋD）患者老年斑周?chē)窠?jīng)元變性和死亡的主要原因。

Aβ（1-42，1-43）是構成老年斑和神經(jīng)纖維纏結的主要成分，出現在A(yíng)D患者的海馬體、大腦皮層和杏仁核處。Aβ（39-43）是形成AD和晚期唐氏綜合癥患者淀粉樣蛋白斑的主要組分。Aβ（1-42）可下調bcl-2且提高bax表達水平。此多肽通過(guò)作用于p75神經(jīng)營(yíng)養素受體來(lái)誘導神經(jīng)死亡。

本品為HFIP處理的人β-淀粉樣蛋白1-42，去除了原合成多肽中預先存在的結構（β-折疊、聚集等），此步處理對于控制聚集研究至關(guān)重要。

產(chǎn)品特性

1）   CAS NO：107761-42-2

2）  同義名：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）   純度：≥95%（HPLC）

6）   外觀(guān)：白色凍干粉末

7）   溶解性：溶于DMSO

8）   單字母序列：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

保存與運輸方法

保存：-20℃避光干燥保存，一年有效。

運輸：冰袋運輸。

注意事項

1）   本品抗衡離子（Counter Ion）是TFA鹽。

2）   本品以?xún)龈煞坌问教峁?，可能因量少不易觀(guān)察到。請直接加溶劑到瓶子使其溶解。

3）   為了您的安全和健康，請穿實(shí)驗服并戴一次性手套操作。

相關(guān)產(chǎn)品

附錄I：不同聚集形態(tài)的合成β-淀粉樣多肽的制備方法（Preparing Synthetic Aβ in Different Aggregation States）【摘自文獻：PMID: 20967580】

圖：制備寡聚體和纖絲狀Aβ1-42的流程匯總圖

1)     用HFIP（六氟異丙醇）處理合成多肽，去除現存的聚合物和β-折疊二級結構，形成單體形式的多肽。（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 DMSO stock by adding 20 μL fresh dry DMSO 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制備物（Unaggregated Aβ Preparation）

13.    Sonicate 5 mM Aβ DMSO 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）寡聚體Aβ制備物（Oligomeric Aβ Preparation）

1.      Start with a tube of freshly resuspended 5 mM Aβ42 in DMSO 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 DMSO, 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）纖絲狀Aβ制備物（Fibrillar Aβ Preparation）

1.      Start with a tube of freshly resuspended 5 mM Aβ42 in DMSO 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 DMSO, 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）淀粉樣斑制備物（Plaque in a Dish” Preparation）

1.      Start with a tube of freshly resuspended 5mM Aβ42 in DMSO 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