DSP交联剂将蛋白质附着在金表面 

 

研究人员经常需要将蛋白质固定在惰性表面上，用于不同的应用，包括分离和纯化ARGET大分子，选择性去除污染物，以及酶催化和化学修饰。固定化酶在金电极上广泛应用于用于检测和分析大分子的亲和生物传感器。

Researchers often need to immobilize proteins on inert surfaces for varied applications including isolation and purification of arget macromolecules, selective removal of contaminants, and enzymatic catalysis and chemical modification. Immobilized enzymes on gold foil electrodes have been used widely in affinity biosensors for the capture and detection of macromolecules for analytical purposes.

化学交联也许是最常用的固定蛋白质在惰性表面上的方法。以下是使用化学交联剂DSP二硫代氨基甲酸酯（琥珀酰亚胺丙酸酯）将蛋白质附着在金表面上的过程。DSP是一种同官能、胺反应的交联剂。在DSP与金表面之间形成的连接是非常稳定的，超过了与玻璃共价硅烷键的强度和稳定性。DSP中的二硫键化学反应迅速地到达金表面，而DSP两端的活性NHS基团对蛋白质中的伯胺基团反应。

Chemical cross-linking is perhaps the most commonly used method for immobilizing proteins on inert surfaces. Following is a protocol for attaching a protein onto a gold surface using the chemical cross-linker dithiobis(succinimidyl propionate) . DSP is a homobifunctional, amine-reactive cross-linker. The linkage formed between DSP and the gold surface is very stable, exceeding the strength and stability of covalent silane bonds with glass. The disulfide linkage in DSP chemisorbs rapidly to gold surfaces, while the active NHS groups on either end of DSP are reactive toward primary amine groups in proteins.

 

产品名称：DSP

简介：Dithiobis(succinimidyl propionate) (DSP) is a homobifunctional crosslinker that is cell membrane permeable. DSP has amine-reactive N-hydroxysuccimide (NHS) esters at both ends of a cleava-ble, 8-atom (12.0 angstrom) spacer arm. DSP contains a reducible disulfide bond in the spacer arm that make it a good choice for various applications.

结构式：

http://s6/bmiddle/003ap78czy7jWvqTRsNd5&690

Molecular Weight: 404.4

CAS #: 57757-57-0

Spacer Length: 12.0 Å

Alternative Names: 3,3′-Dithiodipropionic acid di(N-hydroxysuccinimide ester);

DSP交联剂将蛋白质附着在金表面上

所需材料

• DSP, 4 mg required per ml of DMSO coating solution

• Dimethylsulfoxide (DMSO), sufficient volume to cover and later rinse gold surface

• Protein solution: 1-2 mg/ml protein dissolved in a compatible buffer (pH 7-8, containing at least 50 mM NaCl).

Recommended buffers include PBS, MOPS and HEPES. Avoid buffers containing primary amines,such as Tris and glycine.

推荐的缓冲区包括PBS、MOP和HEPES。避免含有伯胺的缓冲液，如TrIS和甘氨酸。

Note: This procedure can be used to couple other ligands besides proteins that contain primary amines. For molecules that are not soluble in aqueous buffers, coupling can be performed in acetone, isopropyl alcohol or other solvents like DMSO, as long as amines are maintained in an uncharged state by addition of triethylamine or triethanolamine at a concentration equal to or greater than the concentration of DSP.

Procedure

1. Weigh DSP into a tube, and dissolve it in DMSO. Use 4 mg DSP per ml of DMSO necessary to cover the gold surface.

2. Incubate gold foil in dissolved DSP for 30 minutes at room temperature. 3. Rinse gold foil with DMSO and then with water. Gold foil is now activated with NHS groups. Note: Proceed directly to the next step; if protein coupling is not performed immediately, the NHS reactive groups will hydrolyze and protein coupling will be negligible.

3. Immediately add the protein solution to the activated gold foil and incubate for 1-4 hours at room temperature. Note: Incubation for longer time will not adversely affect conjugation efficiency. However, conjugation does not advance significantly after the first 1-2 hours.

4. Rinse the surface with buffer to remove cross-linker by-products (NHS leaving groups) and unconjugated protein.

Note: Free NHS groups (by-products of conjugation and hydrolysis) absorb strongly at 280 nm. Therefore, take care if attempting to determine the conjugation efficiency by measuring the change in protein concentration in solution.

 

应用：

•细胞裂解前的化学交联和免疫沉淀

• “固定”蛋白质相互作用，以确定弱或短暂的蛋白质相互作用

• 固定组织进行免疫染色而不损伤RNA

• 蛋白质交联通过单级反应产生生物偶联物

• 将蛋白质固定在胺涂层表面上

 

 

产品列表：

（一）：Mass Spectrometry Standards - Mass Spec Standards （质朴分析相关试剂）

Leu-Enkephalin

Gonadoliberin

Angiotensin I

Neurotensin

Peptide Calibration Standard Set

（二）：protein crosslinkers （蛋白交联相关试剂）

Protein crosslinkers （蛋白交联相关试剂）：

Heterobifunctional Crosslinkers异型双功能交联剂：

SMCC Crosslinker           CAS Number: 64987-85-5

Sulfo-SMCC Crosslinker      CAS Number: 92921-24-9

Sulfo-SANPAH Crosslinker    CAS Number: 102568-43-4

SMPH Crosslinker

PDPH Crosslinker

MBS Crosslinker             CAS Number: 58626-38-3

Sulfo-MBS Crosslinker

SPDP Crosslinker            CAS Number: 68181-17-9

LC-SPDP Crosslinker  

Sulfo-LC-SPDP Crosslinker

SIA Crosslinker              CAS Number: 39028-27-8

SBA Crosslinker             CAS Number: 42014-51-7

SMPB Crosslinker           CAS Number: 79886-55-8

BMPS Crosslinker           CAS Number: 55750-62-4

EMCS Crosslinker           CAS Number: 55750-63-5

GMBS Crosslinker          CAS Number: 80307-12-6

ANB-NOS Crosslinker        CAS Number: 60117-35-3

Homobifunctional Crosslinkers同型双功能交联剂：

BS3 Crosslinker           CAS Number: 82436-77-9

DSS Crosslinker           CAS Number: 68528-80-3

BS2G Crosslinker

DSG Crosslinker          CAS Number: 79642-50-5

DSP Crosslinker           CAS Number: 57757-57-0

DTSSP Crosslinker        CAS Number: 81069-02-5

Sulfo-EGS Crosslinker     CAS Number: 167410-92-6

EGS Crosslinker          CAS Number: 70539-42-3

DST Crosslinker          CAS Number: 62069-75-4

Zero-Length Crosslinkers - EDC-HCl, Sulfo-NHS & NHS：

EDC-HCl               CAS Number: 25952-53-8

Sulfo-NHS              CAS Number: 106627-54-7

Deuterated Crosslinker BS3-d4

Deuterated Crosslinker BS2G-d4

Deuterated Crosslinker DSS-d4

（三）：Protein modification regents （蛋白修饰相关试剂）

Protein Modification Reagents 蛋白质修饰剂：

SATA Protein Modifier

N-Succinimidyl-S-Acetyl-Thioacetate

CAS Number: 76931-93-6

Sulfo-NHS-Acetate

CAS Number: 152305-87-8

Traut's Reagent  

CAS Number: 4781-83-3

HPG

CAS Number: 24645-80-5

Iodoacetic Acid

CAS Number: 64-69-7

BMPA Modifier

CAS Number: 7423-55-4

（四）：生物素交联试剂

Biotinylation Reagents生物素化试剂

Sulfo-NHS-Biotin     CAS Number: 119616-38-5

NHS-Biotin          CAS Number: 35013-72-0

Sulfo-NHS-LC-Biotin  CAS Number: 127062-22-0

NHS-LC-Biotin       CAS Number: 72040-63-2

Sulfo-NHS-SS-Biotin  CAS Number: 325143-98-4

NHS-SS-Biotin       CAS Number: 142439-92-7

Biotin Hydrazide      CAS Number: 66640-86-6

Biotin-HPDP         CAS Number: 129179-83-5

Iodoacetyl-LC-Biotin   CAS Number: 93285-75-7

（五）Immobilization Resins固定化树脂

Immobilization Resins固定化树脂

Amino-Reactive Resin

Sulfhydryl-Reactive Resin

Carbohydrate-Reactive Resin

Carboxyl-Reactive Resin

Immobilized Pepsin  

Immobilized Chymotrypsin

 

 

以上资料源于西安瑞禧生物科技有限公司

Xi'an ruixi Biological Technology Co;Ltd