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一種可以氨基和巰基反應(yīng)蛋白交聯(lián)劑-SPDP(68181-17-9)
SPDP,CAS #: 68181-17-9 產(chǎn)品可以偶連帶氨基的蛋白和巰基或馬來(lái)酰亞胺的蛋白,偶連帶活性基團(tuán)的蛋白示意圖:
我們可以提供三種不同結(jié)構(gòu)的SPDP異端功能蛋白交聯(lián)劑:SPDP,LC-SPDP,Sulfo-LC-SPDP
其中SPDP,LC-SPDP是脂溶性蛋白交聯(lián)劑,Sulfo-LC-SPDP是水溶性蛋白交聯(lián)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(chǎn)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(jiǎn)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(lián)劑是可分解的,不溶于水,氨和硫醇(巰基)反應(yīng)heterobi功能蛋白交聯(lián)劑。SPDP交聯(lián)試劑的反應(yīng)的moites由N-羥基琥珀酰亞胺(NHS)酯與伯胺反應(yīng)。在另一端的pyridinyldisulfide SPDP,反應(yīng)sulhydryls產(chǎn)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(chǎn)品名稱:Sulfo-LC-SPDP
英文名稱:(sulfosuccinimidyl 6-[3′-(2-pyridyldithio)propionamido]hexanoate)
簡(jiǎn)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(lián)試劑磺可裂解、水溶性氨基和巰基反應(yīng)異型雙功能交聯(lián)劑(巰基)蛋白。
SPDP交聯(lián)劑的反應(yīng)moites磺LC組成的N-羥基(磺基NHS)酯與伯胺反應(yīng)。的LC SPDP交聯(lián)分子pyridinyldisulfide磺化反應(yīng)sulhydryls產(chǎn)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(chǎn)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(lái)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(lái)酰亞胺十一烷酸 N-琥珀酰亞胺酯,CAS#:87981-04-2
2-馬來(lái)酰亞胺基乙酸,CAS#:25021-08-3
11-Maleimidoundecanoic acid(KMUA)CAS#:57079-01-3
Sulfo-EMCS, ε-馬來(lái)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
以上資料源于西安瑞禧生物科技有限公司
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