一種可以氨基和巰基反應蛋白交聯劑-SPDP（68181-17-9）

SPDP，CAS #: 68181-17-9 產品可以偶連帶氨基的蛋白和巰基或馬來酰亞胺的蛋白，偶連帶活性基團的蛋白示意圖:

我們可以提供三種不同結構的SPDP異端功能蛋白交聯劑：SPDP,LC-SPDP,Sulfo-LC-SPDP

其中SPDP,LC-SPDP是脂溶性蛋白交聯劑，Sulfo-LC-SPDP是水溶性蛋白交聯劑。

Procedure for Conjugating Two Proteins, One of Which Contains Sulfhydryls

To conjugate between primary amines on one protein and sulfhydryls (-SH) already present on a second protein, only the first protein must be modified with the SPDP reagent.2 Then, after desalting the SPDP-modified protein to remove reaction byproducts and excess nonreacted reagent, the sulfhydryl-containing protein is added to make the final conjugate (Figure 2, Reaction 1)

Additional Materials Required

Water-miscible organic solvent such as dimethylsulfoxide (DMSO) or

dimethylformamide (DMF); not necessary for Sulfo-LC-SPDP

Phosphate buffered saline with EDTA (PBS-EDTA): 100mM sodium phosphate, 150mM NaCl, 1mM EDTA, 0.02% sodium azide, pH 7.5

Proteins to be conjugated: In the procedure, IgG and β-galactosidase represent the amine-containing and sulfhydrylcontaining protein, respectively.

Procedure

Equilibrate the vial of SPDP Reagent to room temperature before opening. 2.

Immediay before use, prepare a 20mM solution of SPDP Reagent:

Add 25µL of the 20mM SPDP solution to 2-5mg IgG dissolved in 1.0mL of PBS-EDTA.

Incubate for 30 minutes at room temperature.

Equilibrate a desalting column with PBS-EDTA, and buffer exchange the SPDP-modified IgG to remove reaction byproducts and excess nonreacted SPDP reagent

Add 1-3mg of β-galactosidase (~1-3 moles β-gal per mole of IgG) to the IgG solution and incubate the reaction mixture overnight at room temperature

The proteins (IgG and β-galactosidase in this example) are now conjugated.

Additional Information

Please visit the web site for additional information on this product including the following items:

• Tech Tip #1: Attach a protein onto glass, silica or quartz surface using a cleavable crosslinker

• Tech Tip #26: Prepare a reversible and measureable sulfhydryl-reactive affinity column

Protocol For Pyridine-2-Thione Assay to Determine Level of SPDP-modification

1. Dilute 100µL of SPDP-modified and desalted protein to 1mL with PBS.

2. Measure and record the absorbance at 343nm of the protein sample compared to PBS-EDTA blank (test in triplicate)

3. Add 10µL of 15mg/mL DTT to the 1mL SPDP-modified protein sample, mix.

4. After exactly 15 minutes, measure and record the absorbance at 343nm of the reduced sample.

5. Calculate the change in absorbance: ?A343 = (Ave. A343 after DTT) – (Ave. A343 before DTT)

6. Calculate molar ratio of SPDP to Protein using the following equation:

Where the value 8080 reflects the extinction coefficient for pyridine-2-thione at 343 nm: 8.08 x 103 M-1 cm-1

產品名稱：SPDP

英文名稱：N-Succinimidyl 3-(2-Pyridyldithio)propionate

中文名稱：3-(2-吡啶基二硫基)丙酸N-羥基琥珀酰亞胺酯

CAS #: 68181-17-9

簡介：SPDP crosslinker is a cleavable, water insoluble, amino and thiol (sulfhydryl) reactive heterobi-functional protein crosslinker. The SPDP crosslinking reagent's reactive moites consist of an N-hydroxysuccinimide (NHS) ester that reacts with primary amines. On the other end of SPDP, the pyridinyldisulfide reacts with sulhydryls to yield a reversible disulfide bond.

SPDP交聯劑是可分解的，不溶于水，氨和硫醇（巰基）反應heterobi功能蛋白交聯劑。SPDP交聯試劑的反應的moites由N-羥基琥珀酰亞胺（NHS）酯與伯胺反應。在另一端的pyridinyldisulfide SPDP，反應sulhydryls產生可逆的二硫鍵。

Application

3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP) is useful for the preparation of enzyme immunoconjugates and hapten carrier molecule conjugates.

It has been used for antibody conjugation and as a protein cross-linker.

General description

3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP) is a heterobifunctional cross-linking reagent with amine and sulfhydryl reactivity. It is typically coupled initially to molecules containing primary amines by amide bonds buffered at pH 7.5 (6.5-8.5). Second coupling specific for molecules containing free sulfhydryl by thiol-disulfide exchange buffered at pH 8.0 (7.5-8.5). Alternay, this reagent may be used as a protected thiolating reagent following initial coupling and reduction of linker. It incorporates four atom linker.

外觀：白色粉末

Molecular Weight: 312.4

分子式:C12H12N2O4S2

CAS #: 68181-17-9

溶解：DMSO or DMF

Spacer Length: 6.8 Å

Alternative Name: 3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester

純度：98%+

保存：-20℃

：西安瑞禧生物科技有限公司

產品名稱：Sulfo-LC-SPDP

英文名稱：(sulfosuccinimidyl 6-[3′-(2-pyridyldithio)propionamido]hexanoate)

簡介：Sulfo-LC-SPDP crosslinking reagent is a cleavable, water soluble, amino and thiol (sulfhydryl) reactive heterobifunctional protein crosslinker.

Sulfo-LC-SPDP crosslinker's reactive moites consist of an N-hydroxysulfosuccinimide (Sulfo-NHS) ester that reacts with primary amines. The other end of the Sulfo-LC-SPDP crosslinking molecule pyridinyldisulfide reacts with sulhydryls to yield a reversible disulfide bond.

LC SPDP交聯試劑磺可裂解、水溶性氨基和巰基反應異型雙功能交聯劑（巰基）蛋白。

SPDP交聯劑的反應moites磺LC組成的N-羥基（磺基NHS）酯與伯胺反應。的LC SPDP交聯分子pyridinyldisulfide磺化反應sulhydryls產生可逆的二硫鍵的另一端。

Caution

This product is sensitive to moisture. The vial is packaged in a resealable bag with a desiccant to reduce exposure to moisture. After cold storage, equilibrate the vial to room temperature before opening to reduce condensation inside the vial. Make fresh solutions. Storage of stock solutions is not recommended. After use, return the vial to the resealable bag. Close the bag and store the product at the recommended temperature.

General description

The SPDP reagents are a unique group of amine- and sulfhydryl-reactive heterobifunctional crosslinkers. Whether they are used to form amine-to-amine or amine-to-sulfhydryl crosslinks among molecules, the SPDP reagents produce disulfide-containing linkages that can be cleaved later with reducing agents such as dithiothreitol (DTT).

外觀：白色粉末

Molecular Weight: 527.6

分子式：C14H16N3NaO8S3

溶解：水中

Spacer Length: 15.7 Å

Alternative Name: Sulfosuccinimidyl-6-(3’-[2-pyridyldithio]-propionamido) hexanoate

純度：98%+

保存：-20℃

：西安瑞禧生物科技有限公司

產品列表：

DSS        CAS #: 68528-80-3

DTSSP    CAS #: 81069-02-5

EGS       CAS #: 70539-42-3

Sulfo-EGS    

DSSeb    CAS # 23024-29-5

BMPS     CAS #: 55750-62-4

EMCS     CAS #: 55750-63-5

GMBS    CAS #: 80307-12-6

LC-SPDP      

MBS       CAS #: 58626-38-3

PDPH           

SBA       CAS #: 42014-51-7

SIA        CAS #: 39028-27-8

SMCC     CAS #: 64987-85-5

SMPB     CAS #: 79886-55-8

SMPH    

SPDP      CAS #: 68181-17-9

Sulfo-LC-SPDP    

Sulfo-MBS

Sulfo-SANPAH            CAS #: 102568-43-4

Sulfo-SMCC                 CAS #: 92921-24-9

Biotin Hydrazide           CAS #: 66640-86-6

NHS-Biotin                  CAS #: 35013-72-0

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

Sulfo-NHS-Biotin

HPG                            CAS #: 24645-80-5

SATA                           CAS #: 76931-93-6

Sulfo-NHS Acetate        CAS #: 152305-87-8

Biotin-maleimide         

Biotin ethylenediamine hydrobromide  CAS# 216299-38-6

Biotinamidohexanoic acid hydrazide     CAS#: 109276-34-8

CAS#: 89889-52-1

TSAT

BMPH           CAS#: 359436-61-6

EMCH           CAS#: 151038-94-7

KMUH           CAS#: 1593661-78-9

BMOE           CAS: 5132-30-9

BMB              CAS#: 28537-70-4

BMH (bismaleimidohexane)         CAS#:4856-87-5

TMEA

DTME           CAS#:71865-37-7

SATP             CAS#: 84271-78-3

EMCA           CAS: 55750-53-3

AEDP           

Sulfo-NHS，N-羥基琥珀酰亞胺磺酸鈉鹽，CAS #: 106627-54-7

Biotin-ONP，(+)--4-硝基苯酯，CAS#: 33755-53-2

Biotin-PFP，五氟苯酚酯/五氟苯酯，CAS#:120550-35-8

6-(馬來酰亞胺基)己酸琥珀酰亞胺酯，CAS#:55750-63-5

6-Maleimidohexanoic acid，CAS#：55750-53-3

BS3  CAS #: 82436-77-9

BS2G

BSG             CAS #: 79642-50-5

DSP        CAS #: 57757-57-0

N-Maleoyl-β-alanine（MPA），CAS#：7423-55-4

4-Maleimidobutyric acid（MBA），CAS#：57078-98-5

11-馬來酰亞胺十一烷酸 N-琥珀酰亞胺酯，CAS#：87981-04-2

2-馬來酰亞胺基乙酸，CAS#：25021-08-3

11-Maleimidoundecanoic acid（KMUA）CAS#：57079-01-3

Sulfo-EMCS, ε-馬來酰亞胺己酸磺基琥珀酰亞胺酯  CAS#：215312-86-0

AMAS, CAS#：55750-61-3

1,2-Bis(maleimido)ethane, CAS#：5132-30-9

Boc-6-Ahx-OSu, CAS#：51513-80-5

N-Boc-2,2-(ethylenedioxy)diethylamine ,CAS#：153086-78-3

N,N,N,N-Tetramethyl-p-phenylenediamine, CAS#：100-22-1

N-Succinimidyl 6-Biotinamidohexanoate, CAS#：72040-63-2

N-(+)-Biotinyl-6-aminohexanoic acid  CAS#：72040-64-3

CAS#：127062-22-0

DTSSP

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