Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记1mg蛋白 货号5506-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记1mg蛋白

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记1mg蛋白

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记1mg蛋白    货号5506 货号 5506 存储条件 在2-8度冷藏保存, 避免光照
规格 5 Labelings 价格 18288
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

Buccutite 过氧化物酶(HRP)抗体标记试剂盒提供了一种以微观方式标记抗体的便捷方法。蛋白质 – 蛋白质缀合通常用双功能接头(例如常用的SMCC)进行,在每个末端具有不同的反应性以连接两种不同的蛋白质。交联剂的一端(通过NHS酯)与氨基酸赖氨酸和N-末端中发现的胺(-NH2)反应,另一端(通过马来酰亚胺)与氨基酸中发现的硫醇基团(-SH)反应。半胱氨酸。然而,SMCC修饰的蛋白质极不稳定并且通常是自身反应性的,因为蛋白质通常含有胺和硫醇基团,其引起显着量的均聚交联。此外,量化蛋白质上马来酰亚胺基团的数量是相当困难和繁琐的。 Buccutite 过氧化物酶(HRP)抗体偶联试剂盒设计用于直接从蛋白质,肽和含有游离氨基的其他配体制备辣根过氧化物酶(HRP)偶联物。我们的试剂盒中提供的HRP已使用我们专有的接头Buccutite FOL预先活,可直接用于缀合。 Buccutite FOL活化的HRP在极其温和的中性条件下容易与含有Buccutite MTA的分子反应,无需任何催化剂。与常用的SMCC和其他类似技术相比,我们的Buccutite 生物共轭系统更易于使用。它能够以更高的效率和产量实现生物分子的更快和定量缀合。金畔生物是AAT Bioquest的中国代理商,为您提供优质的Buccutite 过氧化物酶(HRP)抗体偶联试剂盒。 

产品说明书

实验方案

操作步骤

1.运行Antibody-Buccutite MTA反应

1.1将抗体溶液直接加入到Buccutite MTA(组分B)的小瓶中,并通过反复移液几次将其充分混合或将小瓶涡旋几秒钟。

1.2将抗体-Buccutite MTA反应混合物在室温下保持30-60分钟。 注意:如果需要,抗体-Buccutite MTA反应混合物可以旋转或摇动更长时间。

 

2.制备抗体-HRP缀合物

2.1通过向Buccutite FOL活化的HRP(组分A)的小瓶中加入50μLddH2O制备Buccutite FOL活化的HRP溶液,通过反复移液几次充分混合或将小瓶涡旋几秒钟。

2.2将整瓶Buccutite FOL活化的HRP溶液混合到纯化的抗体-Buccutite MTA溶液(来自步骤纯化抗体-Buccutite MTA溶液)中,充分混合并在室温下旋转混合物1小时。

2.3抗体-HRP结合物现在可以使用了。 注意:立即使用时,抗体-HRP结合物需要用您选择的缓冲液稀释。 注意:对于长期储存,需要浓缩或冷冻干燥抗体-HRP结合物溶液。

 

3.抗体-HRP共轭物的储存

抗体缀合物应在载体抗体(例如0.1%牛血清白蛋白)存在下以> 0.5mg / mL储存。 当在2mM叠氮化钠存在下储存并避光时,抗体-HRP缀合物溶液可以在4℃下储存两个月而没有显着变化。 对于更长时间的储存,可以将抗体-HRP缀合物冻干并在≤-20℃下储存。

 

参考文献

Chromophore attachment to phycobiliprotein beta-subunits: phycocyanobilin:cysteine-beta84 phycobiliprotein lyase activity of CpeS-like protein from Anabaena Sp. PCC7120
Authors: Zhao KH, Su P, Li J, Tu JM, Zhou M, Bubenzer C, Scheer H.
Journal: J Biol Chem (2006): 8573

Excitation energy transfer from phycobiliprotein to chlorophyll d in intact cells of Acaryochloris marina studied by time- and wavelength-resolved fluorescence spectroscopy
Authors: Petrasek Z, Schmitt FJ, Theiss C, Huyer J, Chen M, Larkum A, Eichler HJ, Kemnitz K, Eckert HJ.
Journal: Photochem Photobiol Sci (2005): 1016

Single-molecule spectroscopy selectively probes donor and acceptor chromophores in the phycobiliprotein allophycocyanin
Authors: Loos D, Cotlet M, De Schryver F, Habuchi S, Hofkens J.
Journal: Biophys J (2004): 2598

Evaluation of Tolypothrix germplasm for phycobiliprotein content
Authors: Prasanna R, Prasanna BM, Mohammadi SA, Singh PK.
Journal: Folia Microbiol (Praha) (2003): 59

Isolation and characterisation of phycobiliprotein rich mutant of cyanobacterium Synechocystis sp
Authors: Prasanna R, Dhar DW, Dominic TK, Tiwari ON, Singh PK.
Journal: Acta Biol Hung (2003): 113

Co-ordinated expression of phycobiliprotein operons in the chromatically adapting cyanobacterium Calothrix PCC 7601: a role for RcaD and RcaG
Authors: Noubir S, Luque I, Ochoa de Alda JA, Perewoska I, Tandeau de Marsac N, Cobley JG, Houmard J.
Journal: Mol Microbiol (2002): 749

Phycobiliprotein genes of the marine photosynthetic prokaryote Prochlorococcus: evidence for rapid evolution of genetic heterogeneity
Authors: Ting CS, Rocap G, King J, Chisholm SW.
Journal: Microbiology (2001): 3171

Novel activity of a phycobiliprotein lyase: both the attachment of phycocyanobilin and the isomerization to phycoviolobilin are catalyzed by the proteins PecE and PecF encoded by the phycoerythrocyanin operon
Authors: Zhao KH, Deng MG, Zheng M, Zhou M, Parbel A, Storf M, Meyer M, Strohmann B, Scheer H.
Journal: FEBS Lett (2000): 9

Phycobiliprotein-Fab conjugates as probes for single particle fluorescence imaging
Authors: Triantafilou K, Triantafilou M, Wilson KM.
Journal: Cytometry (2000): 226

[Phycobiliprotein and fluorescence immunological assay]
Authors: Wu P.
Journal: Sheng Li Ke Xue Jin Zhan (2000): 82

 

相关产品

产品名称 货号
Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记25ug蛋白 Cat#5505
Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记1mg蛋白 Cat#5504

说明书
Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记1mg蛋白.pdf

Buccutite 聚HRP抗体偶联试剂盒 货号5518-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

Buccutite 聚HRP抗体偶联试剂盒

Buccutite 聚HRP抗体偶联试剂盒

Buccutite 聚HRP抗体偶联试剂盒    货号5518 货号 5518 存储条件 在2-8度冷藏保存, 避免光照
规格 1×50 ug Labeling 价格 4836
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

在样品有限或目标分子含量极低的免疫测定中,聚-HRP-抗体结合物被认为具有高的敏感性。 Buccutite 聚HRP抗体偶联试剂盒旨在快速制备聚HRP偶联抗体。该试剂盒提供了可通过我们专有的Buccutite FOL预的poly-HRP探针。用Buccutite MTA(试剂盒中提供)靶向抗体,以得到MTA的抗体。 MTA的抗体与FOL的多HRP的简单混合即可得到所需的多HRP标记的抗体偶联物。该反应在极温和的条件下进行而无需催化剂。用Buccutite Poly-HRP抗体偶联试剂盒制备的poly-HRP抗体偶联物与ELISA,western blotting,免疫组化(IHC)分析中使用的发色,荧光和化学发光HRP底物兼容。它也可以与TSA或我们的Styramide 荧光HRP底物一起使用,以超灵敏地检测低丰度生物靶标。

 

图示

Buccutite 聚HRP抗体偶联试剂盒    货号5518

图1.使用Buccutite Poly-HRP抗体偶联试剂盒(Cat#5518或Cat#5519)制备的polyHRP-山羊抗小鼠IgG偶联物生成直接ELISA曲线。将3倍系列稀释的小鼠IgG包被在96孔板上,并使用标准ELISA方法检测100uL GAM IgG-polyHRP缀合物(100ng / ml)。用TMB底物溶液(Cat#11003)孵育5分钟检测固定的小鼠IgG,并在450 nm处读数。
蓝色:5518或5519试剂盒
红色:GXM IgG PolyHRP(供应商A)
绿色:GXM IgG-HRP(供应商B)

 

参考文献

Detecting cancer metastasis and accompanying protein biomarkers at single cell levels using a 3D-printed microfluidic immunoarray.
Authors: Sharafeldin, Mohamed and Chen, Tianqi and Ozkaya, Gulsum Ucak and Choudhary, Dharamainder and Molinolo, Alfredo A and Gutkind, J Silvio and Rusling, James F
Journal: Biosensors & bioelectronics (2021): 112681

Prostate Cancer Diagnosis in the Clinic Using an 8-Protein Biomarker Panel.
Authors: Jones, Abby L and Dhanapala, Lasangi and Baldo, Thaísa A and Sharafeldin, Mohamed and Krause, Colleen E and Shen, Min and Moghaddam, Shirin and Faria, Ronaldo C and Dey, Dipak K and Watson, R William and Andrawis, Ramez and Lee, Norman H and Rusling, James F
Journal: Analytical chemistry (2020)

Comparison of different methodologies and cryostat versus paraffin sections for chromogenic immunohistochemistry.
Authors: Hira, Vashendriya V V and de Jong, Annique Loncq and Ferro, Klea and Khurshed, Mohammed and Molenaar, Remco J and Van Noorden, Cornelis J F
Journal: Acta histochemica (2019): 125-134

The complementary role of insulin-like growth factor II mRNA-binding protein 3 (IMP3) in diagnosis of Hodgkin’s lymphoma.
Authors: Masoud, Rokia and Ibrahiem, Afaf and Tantawy, Dina and Eldosoky, Ibrahim
Journal: Annals of diagnostic pathology (2019): 64-68

Using magnetic beads and signal amplifiers to produce short and simple immunoassays: Application to MMP-9 detection in plasma samples.
Authors: Ben Ismail, Manel and de la Serna, Erica and Ruiz-Vega, Gisela and García-Berrocoso, Teresa and Montaner, Joan and Zourob, Mohammed and Othmane, Ali and Baldrich, Eva
Journal: Analytica chimica acta (2018): 144-154

Electrochemical immunosensor for sensitive determination of transforming growth factor (TGF) – β1 in urine.
Authors: Sánchez-Tirado, E and Martínez-García, G and González-Cortés, A and Yáñez-Sedeño, P and Pingarrón, J M
Journal: Biosensors & bioelectronics (2017): 9-14

Chemiluminometric Immunosensor for High-Sensitivity Cardiac Troponin I Employing a Polymerized Enzyme Conjugate as a Tracer.
Authors: Lim, Guei-Sam and Seo, Sung-Min and Paek, Sung-Ho and Kim, Seung-Wan and Jeon, Jin-Woo and Kim, Dong-Hyung and Cho, Il-Hoon and Paek, Se-Hwan
Journal: Scientific reports (2015): 14848

Design and construction of novel molecular conjugates for signal amplification (I): conjugation of multiple horseradish peroxidase molecules to immunoglobulin via primary amines on lysine peptide chains.
Authors: Dhawan, Subhash
Journal: Peptides (2002): 2091-8

Design and construction of novel molecular conjugates for signal amplification (II): use of multivalent polystyrene microparticles and lysine peptide chains to generate immunoglobulin-horseradish peroxidase conjugates.
Authors: Dhawan, Subhash
Journal: Peptides (2002): 2099-110

Comparison of primary rabbit kidney and MRC-5 cells and two stain procedures for herpes simplex virus detection by a shell vial centrifugation method.
Authors: Peterson, E M and Hughes, B L and Aarnaes, S L and de la Maza, L M
Journal: Journal of clinical microbiology (1988): 222-4

说明书
Buccutite 聚HRP抗体偶联试剂盒.pdf

Buccutite 聚HRP抗体偶联试剂盒 货号5519-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

Buccutite 聚HRP抗体偶联试剂盒

Buccutite 聚HRP抗体偶联试剂盒

Buccutite 聚HRP抗体偶联试剂盒    货号5519 货号 5519 存储条件 在2-8度冷藏保存, 避免光照
规格 2×50 ug Labelings 价格 6060
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

在样品有限或目标分子含量极低的免疫测定中,聚-HRP-抗体结合物被认为具有高的敏感性。 Buccutite 聚HRP抗体偶联试剂盒旨在快速制备聚HRP偶联抗体。该试剂盒提供了可通过我们专有的Buccutite FOL预的poly-HRP探针。用Buccutite MTA(试剂盒中提供)靶向抗体,以得到MTA的抗体。 MTA的抗体与FOL的多HRP的简单混合即可得到所需的多HRP标记的抗体偶联物。该反应在极温和的条件下进行而无需催化剂。用Buccutite Poly-HRP抗体偶联试剂盒制备的poly-HRP抗体偶联物与ELISA,western blotting,免疫组化(IHC)分析中使用的发色,荧光和化学发光HRP底物兼容。它也可以与TSA或我们的Styramide 荧光HRP底物一起使用,以超灵敏地检测低丰度生物靶标。

 

图示

Buccutite 聚HRP抗体偶联试剂盒    货号5519

图1.使用Buccutite Poly-HRP抗体偶联试剂盒(Cat#5518或Cat#5519)制备的polyHRP-山羊抗小鼠IgG偶联物生成直接ELISA曲线。将3倍系列稀释的小鼠IgG包被在96孔板上,并使用标准ELISA方法检测100uL GAM IgG-polyHRP缀合物(100ng / ml)。用TMB底物溶液(Cat#11003)孵育5分钟检测固定的小鼠IgG,并在450 nm处读数。
蓝色:5518或5519试剂盒
红色:GXM IgG PolyHRP(供应商A)
绿色:GXM IgG-HRP(供应商B)

 

参考文献

Detecting cancer metastasis and accompanying protein biomarkers at single cell levels using a 3D-printed microfluidic immunoarray.
Authors: Sharafeldin, Mohamed and Chen, Tianqi and Ozkaya, Gulsum Ucak and Choudhary, Dharamainder and Molinolo, Alfredo A and Gutkind, J Silvio and Rusling, James F
Journal: Biosensors & bioelectronics (2021): 112681

Prostate Cancer Diagnosis in the Clinic Using an 8-Protein Biomarker Panel.
Authors: Jones, Abby L and Dhanapala, Lasangi and Baldo, Thaísa A and Sharafeldin, Mohamed and Krause, Colleen E and Shen, Min and Moghaddam, Shirin and Faria, Ronaldo C and Dey, Dipak K and Watson, R William and Andrawis, Ramez and Lee, Norman H and Rusling, James F
Journal: Analytical chemistry (2020)

Comparison of different methodologies and cryostat versus paraffin sections for chromogenic immunohistochemistry.
Authors: Hira, Vashendriya V V and de Jong, Annique Loncq and Ferro, Klea and Khurshed, Mohammed and Molenaar, Remco J and Van Noorden, Cornelis J F
Journal: Acta histochemica (2019): 125-134

The complementary role of insulin-like growth factor II mRNA-binding protein 3 (IMP3) in diagnosis of Hodgkin’s lymphoma.
Authors: Masoud, Rokia and Ibrahiem, Afaf and Tantawy, Dina and Eldosoky, Ibrahim
Journal: Annals of diagnostic pathology (2019): 64-68

Using magnetic beads and signal amplifiers to produce short and simple immunoassays: Application to MMP-9 detection in plasma samples.
Authors: Ben Ismail, Manel and de la Serna, Erica and Ruiz-Vega, Gisela and García-Berrocoso, Teresa and Montaner, Joan and Zourob, Mohammed and Othmane, Ali and Baldrich, Eva
Journal: Analytica chimica acta (2018): 144-154

Electrochemical immunosensor for sensitive determination of transforming growth factor (TGF) – β1 in urine.
Authors: Sánchez-Tirado, E and Martínez-García, G and González-Cortés, A and Yáñez-Sedeño, P and Pingarrón, J M
Journal: Biosensors & bioelectronics (2017): 9-14

Chemiluminometric Immunosensor for High-Sensitivity Cardiac Troponin I Employing a Polymerized Enzyme Conjugate as a Tracer.
Authors: Lim, Guei-Sam and Seo, Sung-Min and Paek, Sung-Ho and Kim, Seung-Wan and Jeon, Jin-Woo and Kim, Dong-Hyung and Cho, Il-Hoon and Paek, Se-Hwan
Journal: Scientific reports (2015): 14848

Design and construction of novel molecular conjugates for signal amplification (I): conjugation of multiple horseradish peroxidase molecules to immunoglobulin via primary amines on lysine peptide chains.
Authors: Dhawan, Subhash
Journal: Peptides (2002): 2091-8

Design and construction of novel molecular conjugates for signal amplification (II): use of multivalent polystyrene microparticles and lysine peptide chains to generate immunoglobulin-horseradish peroxidase conjugates.
Authors: Dhawan, Subhash
Journal: Peptides (2002): 2099-110

Comparison of primary rabbit kidney and MRC-5 cells and two stain procedures for herpes simplex virus detection by a shell vial centrifugation method.
Authors: Peterson, E M and Hughes, B L and Aarnaes, S L and de la Maza, L M
Journal: Journal of clinical microbiology (1988): 222-4

说明书
Buccutite 聚HRP抗体偶联试剂盒.pdf

Buccutite AP(碱性磷酸酶)抗体偶联试剂盒*针对标记25 ug蛋白进行了优化* 货号5512-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

Buccutite AP(碱性磷酸酶)抗体偶联试剂盒*针对标记25 ug蛋白进行了优化*

Buccutite AP(碱性磷酸酶)抗体偶联试剂盒*针对标记25 ug蛋白进行了优化*

货号 5512 存储条件 在2-8度冷藏保存, 避免光照
规格 2 Labelings 价格 3612
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

蛋白质-蛋白质缀合交联剂通常使用在每个末端具有不同反应性的双功能接头(例如SMCC)连接两个不同的蛋白质。交联剂的一端与赖氨酸和N端的胺(-NH2)反应(通过NHS酯),另一端与半胱氨酸的硫醇基(-SH)反应(通过马来酰亚胺)。但是,SMCC修饰的蛋白质极不稳定,通常会自反应,因为蛋白质包含多个胺基和硫醇基,会引起大量的交联。另外,定量蛋白质中的马来酰亚胺基团的数量是非常困难的。 Buccutite AP(碱性磷酸酶)抗体偶联试剂盒旨在快速制备具有预活化AP的AP偶联物。该试剂盒针对标记25 ug蛋白进行了优化。 Buccutite FOL活化的AP可以在极其温和的中性条件下与Buccutite MTA修饰的抗体轻松反应,而无需任何催化剂。与常用的SMCC和其他类似技术相比,Buccutite 生物缀合系统更加稳定并且易于使用。它能够以更高的效率和产量更快且定量地偶联生物分子。

 

图示

Buccutite AP(碱性磷酸酶)抗体偶联试剂盒*针对标记25 ug蛋白进行了优化*   货号5512

图1.使用Buccutite AP(碱性磷酸酶)抗体偶联试剂盒制备的ALP-山羊-抗小鼠IgG偶联物检测了小鼠IgG剂量曲线。将3倍系列稀释的小鼠IgG包被在96孔板上,然后按照标准ELISA方法,向每个孔中加入100uL GAM IgG-ALP偶联物(1ug / ml)。在孵育30分钟后,将pNPP底物溶液用于检测固定的小鼠IgG,并在400 nm处读数。

 

参考文献

Direct pulp capping with mineral trioxide aggregate: an immunohistologic comparison with calcium hydroxide in rodents.
Authors: Dammaschke, Till and Stratmann, Udo and Wolff, Philipp and Sagheri, Darius and Schäfer, Edgar
Journal: Journal of endodontics (2010): 814-9

Vitamins C and E inhibit apoptosis of cultured human term placenta trophoblast.
Authors: Tannetta, D S and Sargent, I L and Linton, E A and Redman, C W G
Journal: Placenta (2008): 680-90

Anti-alkaline phosphatase antibody positive myasthenia gravis.
Authors: Konishi, Tetsuro and Ohta, Kiyoe and Shigemoto, Kazuhiro and Ohta, Mitsuhiro
Journal: Journal of the neurological sciences (2007): 89-93

Ectopic expression of alkaline phosphatase in proximal tubular brush border membrane of human renal cell carcinoma.
Authors: Prasad, R and Lambe, S and Kaler, P and Pathania, S and Kumar, S and Attri, S and Singh, S K
Journal: Biochimica et biophysica acta (2005): 240-5

[Identification of antigens of Colombian Giardia duodenalis isolates recognized by total IgG and subclasses].
Authors: Hernández, Jenny Fabiola and Duque, Sofía and Arévalo, Adriana and Guerrero, Rafael and Nicholls, Rubén Santiago
Journal: Biomedica : revista del Instituto Nacional de Salud (2003): 263-73

Primary intrapelvic seminoma in Klinefelter’s syndrome.
Authors: Kurabayashi, A and Furihata, M and Matsumoto, M and Sonobe, H and Ohtsuki, Y and Aki, M and Kuwahara, M
Journal: Pathology international (2001): 624-8

Recombinant adenoviral vector-lipofectAMINE complex for gene transduction into human T lymphocytes.
Authors: Di Nicola, M and Milanesi, M and Magni, M and Bregni, M and Carlo-Stella, C and Longoni, P and Tomanin, R and Ravagnani, F and Scarpa, M and Jordan, C and Gianni, A M
Journal: Human gene therapy (1999): 1875-84

A nonradioactive method for localization of endothelin receptor mRNA in situ.
Authors: McEwan, P E and Valdenaire, O and Sutherland, L and Webb, D J and Gray, G A
Journal: Journal of cardiovascular pharmacology (1998): S443-6

Focus luminescence assay: macroscopically visualized foci of human cytomegalovirus and varicella zoster virus infection.
Authors: Heider, H and Schroeder, C
Journal: Journal of virological methods (1997): 311-6

Histological and immunohistochemical diversities, and proliferative activity and grading in osteosarcomas.
Authors: Hasegawa, T and Hirose, T and Seki, K and Hizawa, K and Ishii, S and Wakabayashi, J
Journal: Cancer detection and prevention (1997): 280-7

说明书
Buccutite AP(碱性磷酸酶)抗体偶联试剂盒*针对标记25 ug蛋白进行了优化*.pdf

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记25ug蛋白 货号5505-AAT Bioquest荧光染料

上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记25ug蛋白

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记25ug蛋白

Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记25ug蛋白     货号5505 货号 5505 存储条件 在2-8度冷藏保存, 避免光照
规格 2 Labelings 价格 2472
Ex (nm) Em (nm)
分子量 溶剂
产品详细介绍

简要概述

Buccutite 过氧化物酶(HRP)抗体标记试剂盒提供了一种以微观方式标记抗体的便捷方法。蛋白质 – 蛋白质缀合通常用双功能接头(例如常用的SMCC)进行,在每个末端具有不同的反应性以连接两种不同的蛋白质。交联剂的一端(通过NHS酯)与氨基酸赖氨酸和N-末端中发现的胺(-NH2)反应,另一端(通过马来酰亚胺)与氨基酸中发现的硫醇基团(-SH)反应。半胱氨酸。然而,SMCC修饰的蛋白质极不稳定并且通常是自身反应性的,因为蛋白质通常含有胺和硫醇基团,其引起显着量的均聚交联。此外,量化蛋白质上马来酰亚胺基团的数量是相当困难和繁琐的。 Buccutite 过氧化物酶(HRP)抗体偶联试剂盒设计用于直接从蛋白质,肽和含有游离氨基的其他配体制备辣根过氧化物酶(HRP)偶联物。我们的试剂盒中提供的HRP已使用我们专有的接头Buccutite FOL预先活,可直接用于缀合。 Buccutite FOL活化的HRP在极其温和的中性条件下容易与含有Buccutite MTA的分子反应,无需任何催化剂。与常用的SMCC和其他类似技术相比,我们的Buccutite 生物共轭系统更易于使用。它能够以更高的效率和产量实现生物分子的更快和定量缀合。金畔生物是AAT Bioquest的中国代理商,为您提供优质的Buccutite 过氧化物酶(HRP)抗体偶联试剂盒。 

产品说明书

实验方案

操作步骤

1.运行Antibody-Buccutite MTA反应

1.1将抗体溶液直接加入到Buccutite MTA(组分B)的小瓶中,并通过反复移液几次将其充分混合或将小瓶涡旋几秒钟。

1.2将抗体-Buccutite MTA反应混合物在室温下保持30-60分钟。 注意:如果需要,抗体-Buccutite MTA反应混合物可以旋转或摇动更长时间。

 

2.制备抗体-HRP缀合物

2.1通过向Buccutite FOL活化的HRP(组分A)的小瓶中加入50μLddH2O制备Buccutite FOL活化的HRP溶液,通过反复移液几次充分混合或将小瓶涡旋几秒钟。

2.2将整瓶Buccutite FOL活化的HRP溶液混合到纯化的抗体-Buccutite MTA溶液(来自步骤纯化抗体-Buccutite MTA溶液)中,充分混合并在室温下旋转混合物1小时。

2.3抗体-HRP结合物现在可以使用了。 注意:立即使用时,抗体-HRP结合物需要用您选择的缓冲液稀释。 注意:对于长期储存,需要浓缩或冷冻干燥抗体-HRP结合物溶液。

 

3.抗体-HRP共轭物的储存

抗体缀合物应在载体抗体(例如0.1%牛血清白蛋白)存在下以> 0.5mg / mL储存。 当在2mM叠氮化钠存在下储存并避光时,抗体-HRP缀合物溶液可以在4℃下储存两个月而没有显着变化。 对于更长时间的储存,可以将抗体-HRP缀合物冻干并在≤-20℃下储存。

 

参考文献

Chromophore attachment to phycobiliprotein beta-subunits: phycocyanobilin:cysteine-beta84 phycobiliprotein lyase activity of CpeS-like protein from Anabaena Sp. PCC7120
Authors: Zhao KH, Su P, Li J, Tu JM, Zhou M, Bubenzer C, Scheer H.
Journal: J Biol Chem (2006): 8573

Excitation energy transfer from phycobiliprotein to chlorophyll d in intact cells of Acaryochloris marina studied by time- and wavelength-resolved fluorescence spectroscopy
Authors: Petrasek Z, Schmitt FJ, Theiss C, Huyer J, Chen M, Larkum A, Eichler HJ, Kemnitz K, Eckert HJ.
Journal: Photochem Photobiol Sci (2005): 1016

Single-molecule spectroscopy selectively probes donor and acceptor chromophores in the phycobiliprotein allophycocyanin
Authors: Loos D, Cotlet M, De Schryver F, Habuchi S, Hofkens J.
Journal: Biophys J (2004): 2598

Evaluation of Tolypothrix germplasm for phycobiliprotein content
Authors: Prasanna R, Prasanna BM, Mohammadi SA, Singh PK.
Journal: Folia Microbiol (Praha) (2003): 59

Isolation and characterisation of phycobiliprotein rich mutant of cyanobacterium Synechocystis sp
Authors: Prasanna R, Dhar DW, Dominic TK, Tiwari ON, Singh PK.
Journal: Acta Biol Hung (2003): 113

Co-ordinated expression of phycobiliprotein operons in the chromatically adapting cyanobacterium Calothrix PCC 7601: a role for RcaD and RcaG
Authors: Noubir S, Luque I, Ochoa de Alda JA, Perewoska I, Tandeau de Marsac N, Cobley JG, Houmard J.
Journal: Mol Microbiol (2002): 749

Phycobiliprotein genes of the marine photosynthetic prokaryote Prochlorococcus: evidence for rapid evolution of genetic heterogeneity
Authors: Ting CS, Rocap G, King J, Chisholm SW.
Journal: Microbiology (2001): 3171

Novel activity of a phycobiliprotein lyase: both the attachment of phycocyanobilin and the isomerization to phycoviolobilin are catalyzed by the proteins PecE and PecF encoded by the phycoerythrocyanin operon
Authors: Zhao KH, Deng MG, Zheng M, Zhou M, Parbel A, Storf M, Meyer M, Strohmann B, Scheer H.
Journal: FEBS Lett (2000): 9

Phycobiliprotein-Fab conjugates as probes for single particle fluorescence imaging
Authors: Triantafilou K, Triantafilou M, Wilson KM.
Journal: Cytometry (2000): 226

[Phycobiliprotein and fluorescence immunological assay]
Authors: Wu P.
Journal: Sheng Li Ke Xue Jin Zhan (2000): 82

 

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说明书
Buccutite 过氧化物酶(HRP)抗体偶联试剂盒 适合标记25ug蛋白 .pdf