Amyloid β Peptide 1-42, Mouse, Rat

大小(xiǎo)鼠β-澱粉樣多(duō)肽1-42

産品信息

【提示1】：點擊此處，見(jiàn)/productView.action?id=10698。

【提示2】：Amyloid β Peptide 1-42受自(zì)身氨基酸序列的影響，非常容易聚集，所以常規合成的多(duō)肽非常難保證完全不含聚集結構。如果客戶要确保得到(dào)單體（不含聚集結構）的對應對肽，可采購我司HPIF處理的Amyloid β Peptide 1-42。信息如下(xià)：

産品描述

Aβ，英文名Amyloid β-protein，中文名β-澱粉樣蛋白(bái)，具神經營養性和神經毒性，由β-澱粉樣前體蛋白(bái)（APP）經β-和γ-分泌酶的蛋白(bái)水(shuǐ)解作用産生(shēng)的含39-43個(gè)氨基酸的多(duō)肽，由細胞分泌，沉澱聚積在細胞基質，此過程不僅與神經元的退行性病變有關，而且能(néng)激活一(yī)系列病理事(shì)件(jiàn)，包括星形膠質細胞和小(xiǎo)膠質細胞的激活、血腦(nǎo)屏障的破壞和微循環變化等，是導緻阿爾茨海默病（AD）患者老年(nián)斑周圍神經元變性和死亡的主要原因。

Aβ（1-42，1-43）是構成老年(nián)斑和神經纖維纏結的主要成分，出現在AD患者的海馬體、大腦(nǎo)皮層和杏仁核處。Aβ（39-43）是形成AD和晚期唐氏綜合症患者澱粉樣蛋白(bái)斑的主要組分。Aβ（1-42）可下(xià)調bcl-2且提高(gāo)bax表達水(shuǐ)平。此多(duō)肽通(tōng)過作用于p75神經營養素受體來誘導神經死亡。

産品特性

1） CAS NO：166090-74-0

2） 同義名：β-Amyloid (1-42), mouse, rat; Amyloid β Protein Fragment 1-42 (mouse, rat);

3） 分子式：C199H307N53O59S

4） 分子量：4417.99 g/mol

5） 純度：≥95%（HPLC）

6） 外觀：白(bái)色凍幹粉

7） 溶解性：溶于DMSO

8） 單字母序列：DAEFGHDSGFEVRHQKLVFFAEDVGSNKGAIIGLMVGGVVIA

9） 三字母序列：Asp-Ala-Glu-Phe-Gly-His-Asp-Ser-Gly-Phe-Glu-Val-Arg-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(guāng)幹燥保存，一(yī)年(nián)有效。

運輸：冰袋運輸。

儲存液制備

1） 将低(dī)溫保存的産品置于室溫回溫至少20min，低(dī)速離心；直接往瓶内加入100% DMSO溶解粉末（0.5mg多(duō)肽加入30μl DMSO足夠）；

2） 之後用1×PBS或者其他合适緩沖液稀釋到(dào)1mg/ml；

3） 室溫低(dī)速漩渦混勻（少于1min）。之後根據單次用量分裝凍存，避免反複凍融。

【注意】：以上(shàng)儲存液制備方法僅做參考，若有其他特殊要求，可根據經驗或參考文獻來進行适宜方法的溶解。

注意事(shì)項

1） 本品以凍幹粉形式提供，可能(néng)因量少不易觀察到(dào)。請直接加溶劑到(dào)瓶子内，漩渦震蕩以确保充分溶解；

2） 為(wèi)了您的安全和健康，請穿實驗服并戴一(yī)次性手套操作。  

相(xiàng)關産品

附錄I：不同聚集形态的合成β-澱粉樣多(duō)肽的制備方法（Preparing Synthetic Aβ in Different Aggregation States）【摘自(zì)文獻：PMID: 20967580】

圖：制備寡聚體和纖絲狀Aβ1-42的流程彙總圖

1)     用HFIP（六氟異丙醇）處理合成多(duō)肽，去除現存的聚合物(wù)和β-折疊二級結構，形成單體形式的多(duō)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(wù)（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(wù)（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(wù)（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）澱粉樣斑制備物(wù)（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