乱序肽 (Phe¹,Ser²)-TRAP-6 (FSLLRN) 可作为 TRAP-6、SFLLRN 的非活性对照。
编号:200390
CAS号:374898-11-0
单字母:H2N-FSLLRN-OH
编号: | 200390 |
中文名称: | 六肽(Phe¹,Ser²)-TRAP-6 |
CAS号: | 374898-11-0 |
单字母: | H2N-FSLLRN-OH |
三字母: | H2N N端氨基:N-terminal amino group。在肽或多肽链中含有游离a-氨基的氨基酸一端。在表示氨基酸序列时,通常将N端放在肽链的左边。 -PheL-苯丙氨酸:phenylalanine。系统命名为(2S)-氨基-3-苯基丙酸。是编码氨基酸。是哺乳动物的必需氨基酸。符号:F,Phe。 -SerL-丝氨酸:serine。系统命名为(2S)-氨基-3-羟基丙酸。是编码氨基酸。因可从蚕丝中获得而得名。符号:S,Ser。在丝原蛋白及某些抗菌素中含有 D-丝氨酸。 -LeuL-亮氨酸:leucine。系统命名为(2S)-氨基-4-甲基戊酸。是编码氨基酸。是哺乳动物的必需氨基酸。符号:L,Leu。 -LeuL-亮氨酸:leucine。系统命名为(2S)-氨基-4-甲基戊酸。是编码氨基酸。是哺乳动物的必需氨基酸。符号:L,Leu。 -ArgL-精氨酸:arginine。系统命名为(2S)-氨基-5-胍基戊酸。在生理条件下带正电荷,为编码氨基酸。是幼小哺乳动物的必需氨基酸。符号:R,Arg。 -Asn天冬酰胺:asparagine。L-天冬酰胺的系统命名为(2S)-氨基-3-氨酰基丙酸,是编码氨基酸。符号:N,Asn。D-天冬酰胺存在于短杆菌肽A分子中。 -OHC端羧基:C-terminal carboxyl group。在肽或多肽链中含有游离羧基的氨基酸一端。在表示氨基酸序列时,通常将C端放在肽链的右边。 |
氨基酸个数: | 6 |
分子式: | C34H56N10O9 |
平均分子量: | 748.87 |
精确分子量: | 748.42 |
等电点(PI): | - |
pH=7.0时的净电荷数: | 1.97 |
平均亲水性: | -0.43333333333333 |
疏水性值: | 0.27 |
消光系数: | - |
来源: | 人工化学合成,仅限科学研究使用,不得用于人体。 |
储存条件: | 负80℃至负20℃ |
标签: | 血液学 |
The scrambled peptide (Phe¹,Ser²)-TRAP-6 (FSLLRN) may serve as an inactive control for TRAP-6, SFLLRN.
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多肽H2N-Phe-Ser-Leu-Leu-Arg-Asn-COOH的合成步骤:
1、合成CTC树脂:称取1.83g CTC Resin(如初始取代度约为0.94mmol/g)和2.06mmol Fmoc-Asn(Trt)-OH于反应器中,加入适量DCM溶解氨基酸(需要注意,此时CTC树脂体积会增大好几倍,避免DCM溶液过少),再加入5.16mmol DIPEA(Mw:129.1,d:0.740g/ml),反应2-3小时后,可不抽滤溶液,直接加入1ml的HPLC级甲醇,封端半小时。依次用DMF洗涤2次,甲醇洗涤1次,DCM洗涤一次,甲醇洗涤一次,DCM洗涤一次,DMF洗涤2次(这里使用甲醇和DCM交替洗涤,是为了更好地去除其他溶质,有利于后续反应)。得到 Fmoc-Asn(Trt)-CTC Resin。结构图如下:
2、脱Fmoc:加3倍树脂体积的20%Pip/DMF溶液,鼓氮气30分钟,然后2倍树脂体积的DMF 洗涤5次。得到 H2N-Asn(Trt)-CTC Resin 。(此步骤脱除Fmoc基团,茚三酮检测为蓝色,Pip为哌啶)。结构图如下:
3、缩合:取5.16mmol Fmoc-Arg(Pbf)-OH 氨基酸,加入到上述树脂里,加适当DMF溶解氨基酸,再依次加入10.32mmol DIPEA,4.9mmol HBTU。反应30分钟后,取小样洗涤,茚三酮检测为无色。用2倍树脂体积的DMF 洗涤3次树脂。(洗涤树脂,去掉残留溶剂,为下一步反应做准备)。得到Fmoc-Arg(Pbf)-Asn(Trt)-CTC Resin。氨基酸:DIPEA:HBTU:树脂=3:6:2.85:1(摩尔比)。结构图如下:
4、依次循环步骤二、步骤三,依次得到
H2N-Arg(Pbf)-Asn(Trt)-CTC Resin
Fmoc-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
H2N-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
Fmoc-Leu-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
H2N-Leu-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
Fmoc-Ser(tBu)-Leu-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
H2N-Ser(tBu)-Leu-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
Fmoc-Phe-Ser(tBu)-Leu-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin
以上中间结构,均可在专肽生物多肽计算器-多肽结构计算器中,一键画出。
最后再经过步骤二得到 H2N-Phe-Ser(tBu)-Leu-Leu-Arg(Pbf)-Asn(Trt)-CTC Resin,结构如下:
5、切割:6倍树脂体积的切割液(或每1g树脂加8ml左右的切割液),摇床摇晃 2小时,过滤掉树脂,用冰无水乙醚沉淀滤液,并用冰无水乙醚洗涤沉淀物3次,最后将沉淀物放真空干燥釜中,常温干燥24小试,得到粗品H2N-Phe-Ser-Leu-Leu-Arg-Asn-COOH。结构图见产品结构图。
切割液选择:1)TFA:H2O=95%:5%、TFA:H2O=97.5%:2.5%
2)TFA:H2O:TIS=95%:2.5%:2.5%
3)三氟乙酸:茴香硫醚:1,2-乙二硫醇:苯酚:水=87.5%:5%:2.5%:2.5%:2.5%
(前两种适合没有容易氧化的氨基酸,例如Trp、Cys、Met。第三种适合几乎所有的序列。)
6、纯化冻干:使用液相色谱纯化,收集目标峰液体,进行冻干,获得蓬松的粉末状固体多肽。不过这时要取小样复测下纯度 是否目标纯度。
7、最后总结:
杭州专肽生物技术有限公司(ALLPEPTIDE https://www.allpeptide.com)主营定制多肽合成业务,提供各类长肽,短肽,环肽,提供各类修饰肽,如:荧光标记修饰(CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等),功能基团修饰肽(叠氮、炔基、DBCO、DOTA、NOTA等),同位素标记肽(N15、C13),订书肽(Stapled Peptide),脂肪酸修饰肽(Pal、Myr、Ste),磷酸化修饰肽(P-Ser、P-Thr、P-Tyr),环肽(酰胺键环肽、一对或者多对二硫键环),生物素标记肽,PEG修饰肽,甲基化修饰肽
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