公司新闻»更多

【Biorbyt实验干货】多肽溶解度指南/ Peptide Solubility Guidelines

发布时间:2021-04-20 17:21 点击次数:

【Biorbyt实验干货】多肽溶解度指南/ Peptide Solubility Guidelines

原创 biorbyt 英国biorbyt 
#实验干货2

Finding the ideal peptide solubility can pose a serious challenge to researchers, since improper solubilization can lead to a loss of peptide or experimental failure. 

找到理想的多肽溶解度可能对研究人员构成严峻挑战,因为不正确的增溶作用可能导致肽丢失或实验失败。

 

While many peptides have good solubility in aqueous solutions, others encounter low solubility and insolubility. This is a particular challenge when peptides have long sequences of hydrophobic amino acids. The guide below provides some suggestions to assist in predicting peptide solubility.

尽管许多肽在水溶液中具有良好的溶解性,但也有部分多肽却具有低溶解性和不溶性。当多肽具有长的疏水性氨基酸序列时,这对其溶解将构成一定挑战。以下指南提供了一些有助于预测多肽溶解度的建议。

 

Before dissolving your peptide, please read the recommendations below and perform a solubility test.

在溶解您的多肽之前,请阅读以下建议并进行溶解度测试。

 

Generally, peptides should first be dissolved in distilled, sterile water, particularly peptides of fewer than five residues. For individual peptides, conditions are chosen for optimum solubility based on the given peptide sequence.

通常,首先应将多肽溶解在蒸馏的无菌水中,尤其是少于五个残基的多肽。对于单个肽,根据给定的多肽序列选择最佳溶解度的条件。

 

· Assign a value of -1 to each acidic residue.

•给每个酸性残基赋值-1。

 

· Assign a value of +1 to each basic residue.

•为每个基本残基分配+1值。

 

· Calculate the overall charge of the peptide.

•计算肽的总电荷。

 

   Acidic Residues

Basic Residues 

Asp (D), Glu (E), C-terminal-COOH

Arg (R), Lys (K), His (H) N-terminal-NH2

 

 

If the overall charge of the peptide is positive, try to dissolve the peptide in water.

如果多肽的总电荷为正,请尝试将多肽溶解在水中。

 

· If the peptide cannot be dissolved, try a 10-30% acetic acid solution.

·如果多肽不能溶解,请尝试使用10-30%的乙酸溶液。

 

· If the peptide will still not dissolve, use TFA (< 50 μl) to solubilize the peptide and dilute to the required concentration.

·如果多肽仍然不能溶解,请使用TFA(<50μl)溶解多肽并稀释至所需浓度。

 

If the overall charge of the peptide is negative, try to dissolve the peptide in water.

如果多肽的总电荷为负,请尝试将多肽溶解在水中。

 

· If the peptide does not dissolve, add NH4OH (< 50 μl) and dilute to the desired concentration.

·如果多肽不溶解,请添加NH4OH(<50μl)并稀释至所需浓度。

 

· If the peptide contains Cysteine, then do not use basic solutions to dissolve it, use the below method.

·如果该多肽含有半胱氨酸,则不要使用碱性溶液来溶解它,请使用以下方法。

 

If the overall charge of the peptide is zero, try to dissolve the peptide in organic solvents.

如果多肽的总电荷为零,请尝试将多肽溶解在有机溶剂中。

 

· First, use acetonitrile, methanol, or isopropanol.

·首先,使用乙腈,甲醇或异丙醇。

 

· For very hydrophobic peptides, try dissolving the peptide in a very small amount of DMSO, and dilute with water to the desired concentration.

·对于疏水性很强的多肽,请尝试将多肽溶解在少量的DMSO中,然后用水稀释至所需浓度。

 

· For Cysteine containing peptides use DMF instead of DMSO.

·对于含半胱氨酸的多肽,请使用DMF代替DMSO。

 

Peptide Dissolving Workflow 多肽溶解流程

 

Ensure you know the appropriate concentration at which the peptide should be dissolved. Please see this link to a great molarity calculator to help you determine the proper concentration. 

确保您知道应溶解多肽的适当浓度。可参考以上摩尔浓度计算方式,以帮助您确定适当的浓度。

 

Some general guidelines include:

一般准则包括:

· Peptides shorter than five residues are usually soluble in water or aqueous buffer, except when the entire sequence consists of hydrophobic amino acids.

·少于五个残基的多肽通常可溶于水或水性缓冲液,除非整个序列由疏水性氨基酸组成。

 

· For peptides that tend to aggregate, add 6 M guanidine-HCl or 8 M urea, then proceed with the necessary dilutions.

·对于易于聚集的多肽,添加6 M盐酸胍或8 M尿素,然后进行必要的稀释。

 

· Hydrophilic peptides containing > 25% charged residues and 25% hydrophobic amino acids are usually soluble in water or aqueous buffers.

·含> 25%带电残基和25%疏水氨基酸的亲水多肽通常可溶于水或水性缓冲液。

 

· Peptides containing 50% and more hydrophobic residues might be insoluble or only partly soluble in aqueous solutions.

·含有50%和更多疏水残基的多肽可能不溶于水溶液或仅部分溶于水溶液。

 

· Warming your solution (< 40°C) may aid in dissolving your peptide.

·加热溶液(<40°C)可能有助于溶解多肽。

 

Please consult literature pertaining to your specific protein as the above are only general guidelines.

请参考与您的特定蛋白质有关的文献,因为以上只是一般指导原则。

 

Peptide Characteristics 多肽特性

Peptide Solubility

Hydrophilic

D,E,H,K,Q,R,S,T, hydroxy-proline, pyro-glutamic acid

Hydrophobic

A,F,I,L,M,P,V,W,Y, alpha-amino butyric acid, beta-amino alanine, norleucine

Peptide Stability

Oxidation in mild conditions

C,M

De-amidation, Dehydration, cyclization to pGlu

N, Q, C-terminal residues, N-terminal Q

Degradation during preparation

M,W

Peptide Charge 

Positive

K,R,H, N-terminus

Negative

D,E,Y, C-terminus


biorbyt拥有各种研究领域的抗体,蛋白和试剂盒,欢迎访问biorbyt全球官方网站(www.biorbyt.com)和中国网站(www.biorbyt.com.cn),或者拨打biorbyt中国办事处热线电话:400-966-3101;027-87663101。 或微信(电话)15307197287在线沟通。