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西安凱新生物科技有限公司
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小分子PEG
DM4PEG m-PEG16-alcoho Pyrene-PEG3-azide N3-PEG9-COOH HO-PEG16-OH N3-PEG4-COOH N3-PEG5-NH2 Ms-PEG3-Ms PEG13 m-PEG10-alcohol mPEG7-COOH m-PEG3-alcohol Tr-PEG9 THP-PEG4 Acid-PEG6-SS-PEG6-acid H2N-PEG8-COOH 2-(t-Butoxycarbonylamido)-1 Dibenzocyclooctyne-acid HO-PEG10-tBu DM1 THP-PEG9 m-PEG8-alcohol m-PEG8-Ms 2-(Biotin-amido) N3-PEG6-CH2CH2COOtBu Mal Tr-PEG3 Fmoc m-PEG9-CH2COOH THP-PEG6 m-PEG7-CH2COOH Propargyl-PEG7-alcohol N3-PEG5-SPA, N3-PEG1-CH2CO2H PEG21 N-Boc-Tris Acid-PEG3-SS-PEG3-acid N3-PEG2-COOH DBCO-S-S-PEG3-biotin m-PEG3-Ms m-PEG6-CH2COOH THP-PEG7 Sulfo-NHS-Acetate Mal-C4-NH-Boc Azido-SS-PEG2-acid N3-PEG10-NHS Tr-PEG6 1,3-bis(carboxyethoxy)-2,2-bis( N3-PEG9-NH2 PEG16 Tr-PEG7 N3-PEG12-COOH m-PEG13-Ms Tos-PEG6-Tos MC-MMAF SPDP m-PEG9-alcohol PEG15 Tetrazine-Acid N3-PEG10-COOH SPDP-C6-NHS ester t-Boc-Cystamine N3-PEG5-OH PEG8-Tos N3-PEG8-OH Ms-PEG8-Ms MC-Val-Cit-PAB-OH N3-PEG36-COOH Acid-PEG2-SS-PEG2-acid Ald-PEG4-azide Sulfo-NHS Auristatin E, THP-PEG12 N3-PEG6-NH2 N3-PEG8-NH2 m-PEG5-CH2COOH m-PEG4-alcohol m-PEG6-Ms PEG12 m-PEG2-COOH PROPARYL-PEG3-ME m-PEG4-CH2COOH N3-PEG8-NHS N3-PEG7-OH N3-PEG4-CH2CH2COOtBu THP-PEG3 PEG5-Ms Azide-SS-biotin Fmoc-PEG4-Ala-Ala-Asn-PAB m-PEG11-alcohol N3-PEG5-CH2CH2COOtBu N3-PEG1-NH2 2-t-Butoxycarbonylamino-1 Tr-PEG4 Fmoc-Val-Cit-PAB-OH, Thiol PEG Tetrazine-PEG TCO-PEG Sugar PEG Propargyl PEG Poly PEG PEG-X-PEG PEG Tosylate PEG Sulfonic acid PEG Phosphonate PEG PFP ester PEG Peptide PEG NHS ester PEG Maleimide PEG azide PEG Aldehyde PEG Acid Non-PEG linker MeNH-PEG m-PEG Hydroxy(OH) PEG Fluorescent PEG DSPE PEG DNP-PEG DBCO PEG Cleavable PEG Bromo PEG Boc-PEG Biotin PEG Benzyl-PEG Aminooxy PEG Amino PEG Alkyne PEG
磷脂
熒光染料
螯合劑
點(diǎn)擊試劑
高分子PEG
交聯(lián)劑
抑制劑
有機(jī)硅
雜環(huán)砌塊
金屬催化劑
多肽
分析試劑
Biotin

Dye maleimides連接帶巰基的蛋白或生物分子

時間:2019/7/10閱讀:401
分享:

Dye maleimides連接帶巰基的蛋白或生物分子

 

馬來酰亞胺活性基團(tuán)的熒光染料是目前和帶有巰基的生物分子比如說多肽、蛋白或者小分子做標(biāo)記或者偶聯(lián)的常用的試劑。

 

Maleimides是一個對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,馬來酰亞胺和巰基會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.

 

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相關(guān)試劑信息:

Cyanine3 maleimide

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.

 

Cyanine5 maleimide

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

 

Cyanine7 maleimide

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.

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