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主营:化学试剂,CCK-8试剂盒,细胞检测,自噬研究,化学标记,生化试剂等
℡ 4000-520-616
℡ 4000-520-616
Dojindo/Peroxidase Labeling Kit-SH/3/LK09
产品编号:LK09
市  场 价:¥0.00
场      地:美国(厂家直采)
产品分类: 蛋白类>多肽>多肽合成>
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:待定
品      牌: Dojindo
公司分类:
Dojindo/Peroxidase Labeling Kit-SH/3/LK09
商品介绍
DescriptionReferencesDataQ & AManualS.D.S

Product DescriptionPeroxidase Labeling Kit-SH is used mainly for the preparation of peroxidase-labeled IgG for enzyme immunoassay (EIA) and for the preparation of peroxidase-labeled antigen for competitive EIA. SH-reactive peroxidase, a component of this kit, can react with the thiol groups of proteins or other molecules (Fig. 1). This kit contains all the necessary reagents for the labeling process, including reducing agent and storage buffer. SH-reactive peroxidase forms a covalent link with the target molecule. Reducing agent can create free thiol groups in the IgG molecule. When peroxidase-labeled IgG is used for EIA, the labeling efficiency of the SH-reactive peroxidase is high enough to eliminate any postlabeling purification process.. If a high-purity conjugate is required after labeling, simply use an affinity column or a gelpermeation column. When labeling small molecules, excess molecules can be removed by using the filtration tubes included in this kit.

Fig. 1 IgG labeling reaction of SH-reactive peroxidase

Precaution♦ The molecular weight of the reduced protein to be labeled with this kit should be greater than 50,000.♦ The molecular weight of the small thiol compound to be labeled with this kit should be smaller than 5,000.♦ IgG or peroxidase-conjugated IgG is always on the membrane of the filtration tube during the labeling process.♦ If the IgG solution contains other proteins with molecular weight larger than 10,000, such as BSA or gelatin, purify the IgG solution prior to labeling peroxidase with this kit. IgG solution can be purified by IgG Purification Kits (not included in this kit).♦ If the IgG solution contains small insoluble materials, centrifuge the solution and use the supernatant for the labeling.

1. K. Inoue, A. Sugiyama, P. C. Reid, Y. Ito, K. Miyauchi, S. Mukai, M. Sagara, K. Miyamoto, H. Satoh, I. Kohno, T. Kurata, H. Ota, A. Mantovani, T. Hamakubo, H. Daida and T. Kodama, “Establishment of a High Sensitivity Plasma Assay for Human Pentraxin3 as a Marker for Unstable Angina Pectoris”, Arterioscler. Thromb. Vasc. Biol., 2007, 27, 161.2. N. Esaki, Y. Ohkawa, N. Hashimoto, Y. Tsuda, Y. Ohmi, R. H. Bhuiyan, N. Kotani, K. Honke, A. Enomoto, M. Takahashi, K. Furukawa, and K. Furukawa, “ASC amino acid transporter 2, defined by enzyme-mediated activation of radical sources, enhances malignancy of GD2-positive small-cell lung cancer.”, Cancer Sci.., 2018, 109, (1), 141.3. G.W.Zhanga, S.J.Lai, Y.Yoshimura, and N.Isobe, “Messenger RNA expression and immunolocalization of psoriasin in the goat mammary gland and its milk concentration after an intramammary infusion of lipopolysaccharide”, Vet. J.., 2014, 202, (1), 89.4. G-W. Zhang, S-J. Lai, Y. Yoshimura, and N. Isobe, “Expression of cathelicidins mRNA in the goat mammary gland and effect of the intramammary infusion of lipopolysaccharide on milk cathelicidin-2 concentration”, Vet. Microbiol.., 2014, 170, (1-2), 125.5. H. Tateno, S. Saito, K. Hiemori, K. Kiyoi, K. Hasehira, M. Toyoda, Y. Onuma, Y. Ito, H. Akutsu, and J. Hirabayashi, “α2–6 sialylation is a marker of the differentiation potential of human mesenchymal stem cells”, Glycobiology., 2016, 26, (12), 1328.6. K. Iizumi, H. Kawasaki, A. Shigenaga, M. Tominaga, A. Otsu, A. Kamo, Y. Kamata, K. Takamori, and F. Yamakura, “Tryptophan nitration of immunoglobulin light chain as a new possible biomarker for atopic dermatitis”, J Clin Biochem Nutr., 2018, 63, (3), 197.7. K. Morioka, K. Fukai, K. Yoshida, R. Yamazoe, H. Onozato, S. Ohashi, T. Tsuda, and K. Sakamoto, “Foot-and-Mouth Disease Virus Antigen Detection Enzyme-Linked Immunosorbent Assay Using Multiserotype-Reactive Monoclonal Antibodies”, J. Clin. Microbiol.., 2009, 47, (11), 3663.8. M. Watanabe, I. Takemasa, N. Kaneko, Y. Yokoyama, E. Matsuo, S. Iwasa, M. Mori, N. Matsuura, M. Monden, and O. Nishimura, “Clinical significance of circulating galectins as colorectal cancer markers”, Oncol. Rep.., 2011, 25, (5), 1217.9. M. Yasunaga, S. Saijou, S. Hanaoka, T. Anzai, R. Tsumura, and Y. Matsumura, “Significant antitumor effect of an antibody against TMEM180, a new colorectal cancer‐specific molecule”, Cancer Sci.., 2019, 110, (2), 761.10. N. Esaki, Y. Ohkawa, N. Hashimoto, Y. Tsuda, Y. Ohmi, R. H. Bhuiyan, N. Kotani, K. Honke, A. Enomoto, M. Takahashi, K. Furukawa, and K. Furukawa, “ASC amino acid transporter 2, defined by enzyme‐mediated activation of radical sources, enhances malignancy of GD2‐positive small‐cell lung cancer”, Cancer Sci.., 2018, 109, (1), 141.11. N. Hashimoto, K. Hamamura, N. Kotani, K. Furukawa, K. Kaneko, K. Honke, and K. Furukawa, “Proteomic analysis of ganglioside‐associated membrane molecules: Substantial basis for molecular clustering”, Proteomics., 2012, 12, (21), 3154.12. N. Kotani, Y. Ida, T. Nakano, I. Sato, R. Kuwahara, A. Yamaguchi, M. Tomita, K. Honke, and T. Murakoshi, “Tumor-dependent secretion of close homolog of L1 results in elevation of its circulating level in mouse model for human lung tumor”, Biochem. Biophys. Res. Commun.., 2018, 501, (4), 982.13. R. Yamashita, N. Kotani, Y. Ishiura, S. Higashiyama, and K. Honke, “Spatiotemporally-regulated interaction between β1 integrin and ErbB4 that is involved in fibronectin-dependent cell migration”, J. Biol. Chem.., 2011, 149, (3), 347.14. T. Noro, E. Oishi, T. Kaneshige, K. Yaguchi, K. Amimoto, and M. Shimizu, “Identification and characterization of haemagglutinin epitopes of Avibacterium paragallinarum serovar C”, Vet. Microbiol.., 2008, 131, (3-4), 406.

Sandwich ELISA

Fig. 2 Sandwich ELISA of human TNF-α detection

Plate: 2 μg/ml anti-human TNF-aantibody (rabbit, polyclonal)-coated high binding platerecombinant human TNF-a: 0-1000 pg/ml PBSTPeroxidase-conjugated anti-human TNF-aantibody: Prepared by Peroxidase Labeling Kit-SH.1μg/mlPBST+blocking reagentSubstrate: TMB peroxidase substrate

References1. K. Inoue, A. Sugiyama, P. C. Reid, Y, Ito, K. Miyauchi, S. Mukai, M. Sagara, K. Miyamoto, H. Satoh, I. Kohno, T. Kurata, H. Ota, A. Mantovani, T. Hamakubo, H. Daida and T. Kodama, Establishment of a High Sensitivity Plasma Assay for Human Pentraxin3 as a Marker for Unstable Angina Pectoris, Arterioscler. Thromb. Vasc. Biol., 2007, 27, 161

Can I use this kit for F(ab")2?

Yes, please follow the labeling protocol for IgG. The recovery of the conjugate should be over 80%.

Can I use this kit for other proteins or peptides?

Yes, if the molecular weight of the reduced form is greater than 50,000 or less than 5,000, and it has a reactive SH group, or a disulfide group that can be reduced without losing activity. If the molecular weight is greater than 50,000, follow the labeling protocol for IgG, and use 0.5-1 nmol of sample protein for LK09-10. If the molecular weight is less than 5,000, follow the labeling protocol for small molecules. If the molecular weight is between 5,000 and 50,000, contact our customer service at info@dojindo.com or 1-877-987-2667 for more information.

Can I use this kit to label oligopeptides or oligonucleotides?

Yes, if the molecular weights of the oligonucleotide or the oligopeptide are less than 5,000 and they have at least one SH group. Follow the labeling protocol for small molecule.

What is the minimum amount of IgG that can be labeled with LK09-10?

The minimum amount is 50 μg. There is no significant difference in sensitivity and background between 50 μg and 200 μg of IgG. However, even 10 μg IgG can be labeled using 1/5 volume of SH-reactive peroxidase solution at Step 8.

How many peroxidase molecules per reduced IgG are introduced?

The average number of peroxidase molecule per reduced IgG is 1 to 2.

Do I have to use a Filtration tube prior to labeling the protein?

If the protein solution does not contain small molecules with reactive SH groups and the concentration of the protein is 10 mg per ml, or about 70 μM, there is no need to use the Filtration tube. Just mix the sample solution with Solution B and add the mixture to a vial of the SH-reactive peroxidase.

Do I have to use Storage buffer included with the kit?

No, you don’t have to use Storage buffer from the kit. You can choose any kind of buffer appropriate for your experiment. However, the Storage buffer helps to increase the stability of the peroxidase conjugate.

My sample contains small insoluble material. What should I do?

Spin the sample and use the supernatant for labeling.

Does unconjugated SH reactive peroxidase still have a reactive maleimide after the labeling reaction to IgG?

No. Nearly 100% of SH reactive peroxidase is used for the IgG labeling or the small molecule labeling.

Does Storage buffer contain animal products or polymers?

No, Storage buffer does not contain any animal products, polymers, or heavy metal ions.

Related Categories Labeling Chemistry

品牌介绍

Dojindo Molecular Technologies,Inc.是美国领先的生命科学研究专用试剂盒和化学品分销商。我们通过提供优质的支持和创新产品,不断扩大客户的满意度,以扩大生命科学研究和开发领域。

我们的公司文化反映出“ Jin”(仁)字符背后的含义,即尊重和关爱他人。当我们公司执行上述任务时,“ Jin”精神始终处于我们思想的最前沿。  


我们如何开始

1996年

作为日本Dojindo实验室的子公司,Dojindo Molecular Technologies,Inc.在马里兰州盖瑟斯堡开业,这是新产品开发的主要设施。开发了诸如DNA损伤定量试剂盒SOD检测试剂盒WST标记试剂盒系列之类的产品,为全球客户提供现成的检测试剂盒。


2000

Dojindo Molecular Technologies,Inc.成为了另一个销售地点,将Dojindo产品带到了北美地区的研究专业人员手中。


2008年

我们搬迁至马里兰州罗克维尔,仅专注于北美的销售和营销活动。我们目前的位置非常接近美国国立卫生研究院(NIH),美国国家癌症研究所(NCI),FDA,约翰·霍普金斯大学的研究人员。


我们不仅向周边地区,而且向我们在美国,加拿大和南美的客户提供日本优质的客户支持和创新产品。

关于同人堂实验室

Dojindo Laboratories由螯合化学领域的著名研究人员上野敬平教授创立。同人堂实验室是1951年第一家将EDTA(乙二胺四乙酸)商业化的日本公司。自那以后,同人堂实验室一直在生产用于支持科学研究进展的试剂。Dojindo实验室的任务是开发用于生命科学研究的创新工具。Dojindo实验室致力于通过科学发现和开发新药物为改善生活质量做出贡献。


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