馬來(lái)酰亞胺染料連接帶巰基的蛋白或生物分子
馬來(lái)酰亞胺染料連接帶巰基的蛋白或生物分子
馬來(lái)酰亞胺活性基團(tuán)的熒光染料是目前和帶有巰基的生物分子比如說(shuō)多肽、蛋白或者小分子做標(biāo)記或者偶聯(lián)的最常用的試劑。
maleimides是一個(gè)對(duì)thiols有高選擇性的親電化合物，在自然界中基本上不存在maleimides基團(tuán)，而有豐富的thiols存在，馬來(lái)酰亞胺和巰基會(huì)作為半胱氨酸殘基在多肽和蛋白中相遇，雖然天然的dna中不含有巰基，但是人工合成的寡核苷酸是非常容易制備出來(lái)的。
cy3 maleimides , cy5 maleimides , cy7 maleimides標(biāo)記帶有sh的多肽、蛋白還有生物小分子的具體實(shí)驗(yàn)方法和實(shí)驗(yàn)步驟。
1. dissolve the protein or other moleculecontaining thiol to be labeled in degassed buffer (pbs, tris, hepes are good,although others buffers containing no thiols can be used) at ph 7-7.5 inplastic vial. buffer can be degassed by applying vacuum on it for severalminutes, or by bubbling through inert gas (nitrogen, argon, or helium). forproteins, good concentration is between 1-10 mg/ml.
2. add an excess of tcep(tris-carboxyethylphosphine) reagent to reduce disulfide bonds, flush withinert gas, and close. 100x molar excess of tcep is fine. keep the mixture for20 minutes at room temperature.
3. dissolve maleimide in dmso or fresh dmf(1-10 mg in 100 ul).
4. add dye solution to thiol solution (20xfold excess of dye), flush vial with inert gas, and close tightly. 5. mixthoroughly, and keep overnight at room temperature, or 4 celsius. 6. purify bygel filteration, hplc, fplc, or electrophoresis
for maleimides with poor aqueoussolubility, like most dye maleimides, we recommend use of cosolvent (dmf ordmso). maleimides with good aqueous solubility (like sulfo-cy maleimides) canbe dissolved in water. if precipitation occurs, increase content of organicco-solvent in the mixture to achieve better labeling.
相關(guān)試劑信息：
cyanine3maleimide
appearance:
red powder
molecular weight:
615.20
molecular formula:
c36h43cln4o3
solubility:
soluble in organic solvents (dmf, dmso, dichloromethane), insoluble in water
quality control:
nmr 1h (95%) and 13c, tlc, functional testing
storage conditions:
storage: 24 months at -20°c in the dark.
cyanine5maleimide
appearance:
dark blue powder
molecular weight:
641.24
molecular formula:
c38h45cln4o3
solubility:
soluble in organic solvents (dmf, dmso, dichloromethane), insoluble in water
quality control:
nmr 1h (95%) and 13c, tlc, functional testing
storage conditions:
storage: 24 months at -20°c in the dark
cyanine7maleimide
appearance:
green powder
molecular weight:
707.34
molecular formula:
c43h51cln4o3
solubility:
soluble in organic solvents (dmso, dmf, dichloromethane), low solubility in water
quality control:
nmr 1h (95%) and 13c, tlc, functional testing
storage conditions:
storage: 24 months at -20°c in the dark.
以上資料源于西安瑞禧生物科技有限公司
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馬來(lái)酰亞胺活性基團(tuán)的熒光染料是目前和帶有巰基的生物分子比如說(shuō)多肽、蛋白或者小分子做標(biāo)記或者偶聯(lián)的最常用的試劑。
maleimides是一個(gè)對(duì)thiols有高選擇性的親電化合物，在自然界中基本上不存在maleimides基團(tuán)，而有豐富的thiols存在，馬來(lái)酰亞胺和巰基會(huì)作為半胱氨酸殘基在多肽和蛋白中相遇，雖然天然的dna中不含有巰基，但是人工合成的寡核苷酸是非常容易制備出來(lái)的。
cy3 maleimides , cy5 maleimides , cy7 maleimides標(biāo)記帶有sh的多肽、蛋白還有生物小分子的具體實(shí)驗(yàn)方法和實(shí)驗(yàn)步驟。
1. dissolve the protein or other moleculecontaining thiol to be labeled in degassed buffer (pbs, tris, hepes are good,although others buffers containing no thiols can be used) at ph 7-7.5 inplastic vial. buffer can be degassed by applying vacuum on it for severalminutes, or by bubbling through inert gas (nitrogen, argon, or helium). forproteins, good concentration is between 1-10 mg/ml.
2. add an excess of tcep(tris-carboxyethylphosphine) reagent to reduce disulfide bonds, flush withinert gas, and close. 100x molar excess of tcep is fine. keep the mixture for20 minutes at room temperature.
3. dissolve maleimide in dmso or fresh dmf(1-10 mg in 100 ul).
4. add dye solution to thiol solution (20xfold excess of dye), flush vial with inert gas, and close tightly. 5. mixthoroughly, and keep overnight at room temperature, or 4 celsius. 6. purify bygel filteration, hplc, fplc, or electrophoresis
for maleimides with poor aqueoussolubility, like most dye maleimides, we recommend use of cosolvent (dmf ordmso). maleimides with good aqueous solubility (like sulfo-cy maleimides) canbe dissolved in water. if precipitation occurs, increase content of organicco-solvent in the mixture to achieve better labeling.
相關(guān)試劑信息：
cyanine3maleimide
appearance:
red powder
molecular weight:
615.20
molecular formula:
c36h43cln4o3
solubility:
soluble in organic solvents (dmf, dmso, dichloromethane), insoluble in water
quality control:
nmr 1h (95%) and 13c, tlc, functional testing
storage conditions:
storage: 24 months at -20°c in the dark.
cyanine5maleimide
appearance:
dark blue powder
molecular weight:
641.24
molecular formula:
c38h45cln4o3
solubility:
soluble in organic solvents (dmf, dmso, dichloromethane), insoluble in water
quality control:
nmr 1h (95%) and 13c, tlc, functional testing
storage conditions:
storage: 24 months at -20°c in the dark
cyanine7maleimide
appearance:
green powder
molecular weight:
707.34
molecular formula:
c43h51cln4o3
solubility:
soluble in organic solvents (dmso, dmf, dichloromethane), low solubility in water
quality control:
nmr 1h (95%) and 13c, tlc, functional testing
storage conditions:
storage: 24 months at -20°c in the dark.
以上資料源于西安瑞禧生物科技有限公司
東城區(qū)附近裝飾工程公司更受歡迎
養(yǎng)馬護(hù)欄，內(nèi)蒙古草原圍欄，PVC美式柵欄，馬場(chǎng)防護(hù)
寶寶睡眠不足怎么辦
城隍廟展柜制作哪家好|展柜制作
氣動(dòng)單軌吊 礦用運(yùn)輸單軌吊
馬來(lái)酰亞胺染料連接帶巰基的蛋白或生物分子
喜大普奔！月經(jīng)假、崗位補(bǔ)貼、二胎補(bǔ)貼！更多利好政策源源不斷！
預(yù)制聚氨酯發(fā)泡保溫管價(jià)格
供應(yīng)粉末尿素 化妝品級(jí)別 高純度高保濕性能
到底我們的豬場(chǎng)該做哪些疫苗？絕對(duì)長(zhǎng)知識(shí)！
汽車?yán)走_(dá)芯片商機(jī)巨大 中國(guó)還需三五年迎來(lái)爆發(fā)期
復(fù)聯(lián)四：浩克覺(jué)醒后不僅穿上了黑背心，還帶上了機(jī)械手套吊打滅霸
供應(yīng)品牌 蒙娜麗莎 衛(wèi)浴座便器/馬桶
一分鐘解讀為什么不選平臺(tái)電商
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供應(yīng)鋁粉專用振動(dòng)篩-防爆超聲波振動(dòng)篩（圖）
北京主持人培訓(xùn)哪家比較好，行業(yè)領(lǐng)先
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A042不銹鋼焊條
河南煉油設(shè)備生產(chǎn)廠家