中文原理介紹：
寡核苷酸可以通過硫醇-馬來酰亞胺和腙偶聯(lián)方法與抗體/蛋白質偶聯(lián)。硫醇-馬來酰亞胺方法通常使用雙功能接頭（例如 SMCC）進行。然而，SMCC修飾的蛋白質極不穩(wěn)定，并且經常自反應，因為它通常含有胺和硫醇基團，導致大量的同源交聯(lián)。當存在其他硫醇基團時，共軛鍵可交換。在腙偶聯(lián)方法中，共軛反應基于肼和醛之間的反應。盡管使用了催化劑，但這種方法的收率很低。分子的大小和官能團的不可接近性導致產量低。此外，聯(lián)動穩(wěn)定性也是一個問題。
BroadPharm（國內總代理：靶點科技）的MagicLink™技術通過結合PEG和專有的交聯(lián)化學解決了所有問題。MagicLink™ 化學試劑盒和試劑盒根本不需要 TCEP、DTT 或催化劑！
首先，抗體與LINK NHS酯反應，將LINK引入抗體中。此外，胺修飾的寡核苷酸與MAGIC NHS酯反應，將MAGIC引入寡核苷酸結構中。其次，在純化兩個反應產物后，它們立即相互反應，產生具有穩(wěn)定環(huán)連接的抗體-寡核苷酸偶聯(lián)物。
- 新穎性：第三代蛋白質交聯(lián)技術
- 產率：偶聯(lián)效率在95-100%左右
- 速率：即時反應;整個過程不到1小時
- 鍵穩(wěn)定性：市場上穩(wěn)定的共價鍵偶聯(lián)物
- 一致性：批次間重現(xiàn)性好
- 靈活：以所需的比例使用您自己的寡核苷酸和抗體
- 保質期：在室溫下長時間、穩(wěn)定的凍干船
- 抗體選擇：任何蛋白質/抗體、抗體片段
- 寡核苷酸選擇：10~120個堿基的單鏈寡核苷酸，最多80個堿基對的雙鏈寡核苷酸; 連接化學在 5 端和 3 端都有效
- 無需偶聯(lián)后凈化步驟
- 包括陽性對照抗體和寡核苷酸，可確認方案成功
該試劑盒還可用于將寡核苷酸與堿性磷酸酶（AP）、辣根過氧化物酶（HRP）、牛血清白蛋白（BSA）、大豆過氧化物酶（SBP）、青霉素酶（PNC）、鏈霉親和素及其偶聯(lián)物、α-D-半乳糖苷酶、葡萄糖-t-磷酸脫氫酶等偶聯(lián)。
這種寡核苷酸-酶偶聯(lián)物（“寡酶"）可用于多種雜交和檢測形式，包括斑點印跡、南方/北方印跡、原位和溶液雜交/捕獲方案。
原理示意圖：
英文原理介紹：
Oligonucleotide can be conjugated to antibody/protein by thiol-maleimide and hydrazone coupling approaches. The thiol-maleimide approach is commonly performed with a bifunctional linker (such as SMCC). However, SMCC-modified protein is extremely unstable and often self-reactive since it often contains both amine and thiol groups that cause a significant amount of homo-crosslinking. The conjugation linkage is exchangeable when other thiol groups are present. In the hydrazone coupling approach, the conjugation reaction is based on a reaction between hydrazine and aldehyde. This method is low yielding although a catalyst is used. The size of the molecule and inaccessibility of the functional group contribute to low yield. In addition, linkage stability is an issue.
BroadPharm's MagicLink™ technology solves all the problems by combining PEG and proprietary crosslinking chemistry. MagicLink™ chemistry and kit do not need TCEP, DTT, or catalysts at all!
First, antibody reacts with LINK NHS ester to introduce LINK into the antibody. Also, amine modified oligonucleotide reacts with MAGIC NHS ester to introduce MAGIC into the oligonucleotide structure. Second, after purification of both reactive partners, they instantly react with each other yielding an antibody-oligonucleotide conjugate with a stable ring linkage.
- Novelty: 3rd generation protein crosslink technology
- Yield: conjugation efficiency around 95-100%
- Rate: instant reaction; the whole process less than 1 hour
- Linkage stability: most stable covalent bonded conjugates on the market
- Consistence: good reproducibility from batch to batch
- Flexible: use your own oligo and antibody at your desired ratio
- Shelf life: long, stable freeze dried ship at ambient temperature
- Antibody selection: any proteins/antibody, antibody fragment
- Oligo selection: single strand oligos of 10 120 bases, and double strand oligos up to 80 base pairs;
Linking chemistry works at both 5 and 3 end
- Post-conjugation clean-up step not necessary
- Positive control antibody and oligo included to enable confirmation of protocol success
The kit can also be used to conjugate oligonucleotides to alkaline phosphatase (AP), horseradish peroxidase (HRP), bovine serum albumin (BSA), soybean peroxidase (SBP), penicillinase (PNC), streptavidin and its conjugates, alpha-D-galactosidase, glucose-t-phosphate dehydrogenase, etc.
Such oligonucleotide-enzyme conjugates ("oligozymes") can be used in a variety of hybridization and detection formats, including dot blots, Southern/Northern blots, in situ and solution hybridization/capture schemes.
應用舉例：
訂購信息：
名稱：MagicLink™ Oligo Antibody Conjugation Kit
貨號：BP-50004
規(guī)格：3*100ug
總代理：靶點科技
