上海熹垣譜析（上海）生物科技有限公司優(yōu)勢供應(yīng)：DNA Unwinding Assay Kit
2019/10/14 13:36:54
該試劑盒的設(shè)計目的是研究是否有一種化合物插入到DNA雙螺旋結(jié)構(gòu)中，或結(jié)合在溝槽中，導(dǎo)致DNA解開。這是許多dna修飾酶抑制劑的特征，如拓撲異構(gòu)酶抑制劑m-amsacrine。插入器包含平面的，通常是多環(huán)芳香族結(jié)構(gòu)，可以插入到雙螺旋DNA分子的堿基之間。能夠插入DNA或在凹槽中結(jié)合的化合物可以導(dǎo)致DNA的局部解繞，從而導(dǎo)致DNA扭曲的減少。如果一個DNA分子在這種化合物的存在下被切割并重新連接(例如通過拓撲異構(gòu)酶)，那么結(jié)果就是松弛的、未纏繞的DNA;去除酶和化合物后，它會轉(zhuǎn)化成超螺旋DNA。在這些條件下形成的超螺旋DNA表明是插入物。這是DNA解開實驗的基礎(chǔ)。超螺旋(或放松)質(zhì)粒DNA與測試化合物進行短暫孵育，然后由小麥胚芽topo i進行放松。然后提取酶和藥物，凝膠電泳分析質(zhì)粒。超螺旋狀構(gòu)造表明該化合物是一種插入劑或溝槽粘合劑。如果化合物不是插入物，那么產(chǎn)物將是松弛質(zhì)粒。應(yīng)使用放松質(zhì)粒進行對照反應(yīng)，以證明該化合物不是簡單地作為topo I的抑制劑，從而檢查檢測結(jié)果。該試劑盒(產(chǎn)品代碼DUKSR001)包含所有您需要做的分析，包括:-超螺旋pBR322(50μg),放松pBR322(25μg),小麥胚芽威尼斯平底漁船我(250 U),分析緩沖(1毫升)和稀釋緩沖(1毫升)。我們還提供工具包，其中包含topo I和緩沖區(qū)，但只有(i) pBR322(產(chǎn)品代碼DUKR002)的寬松形式。這將顯示一個化合物是否為插入物，但如果它也抑制topo I(即可能出現(xiàn)假陰性)，則不會顯示(ii) pBR322的超卷形式(產(chǎn)品代碼DUKS003)。這將顯示化合物是否為插入劑，但如果它是topo I的抑制劑，則會得到相同的結(jié)果(即可能出現(xiàn)假陽性)。如下圖所示:
This kit is designed to investigate whether a compound intercalates into the DNA double-helix, or binds in the groove, leading to unwinding of the DNA. This is a characteristic of a number of inhibitors of DNA-modifying enzymes such as the topoisomerase inhibitor m-amsacrine.
Intercalators contain planar, normally polycyclic, aromatic structures which can insert between the bases of the double-helical DNA molecule. Compounds which are able to intercalate into DNA or bind in the groove can lead to local unwinding of the DNA leading to a decrease in the twist of the DNA. If a DNA molecule is nicked and rejoined (for example by a topoisomerase) in the presence of such a compound, then the result is relaxed, underwound DNA; on removal of the enzyme and compound this converts to supercoiled DNA. The supercoiled DNA formed in these conditions is indicative of an intercalator.
This is the basis of the DNA unwinding assay. Supercoiled (or relaxed) plasmid DNA is incubated briefly with the test compound prior to relaxation by the wheat germ topo I. The enzyme and drug are then removed by extraction and the plasmid analysed by gel electrophoresis. Supercoiled topoiosmers indicate the compound is an intercalator or groove binder. If the compound is not an intercalator then the product will be relaxed plasmid.
The assay should be checked by performing a control reaction using relaxed plasmid to show that the compound is not simply acting as an inhibitor of the topo I.
The kit (product code DUKSR001) contains all you need to do the assay including:-
Supercoiled pBR322 (50 μg), relaxed pBR322 (25 μg), wheat germ topo I (250 U), Assay Buffer (1 ml) and Dilution Buffer (1 ml).
We also supply kits which contain topo I and buffers but only:-
(i) the relaxed form of pBR322 (product code DUKR002). This will show if a compound is an intercalator but not if it also inhibits the topo I (i.e. false-negatives are possible)
(ii) the supercoiled form of pBR322 (product code DUKS003). This will show if a compound is an intercalator but the same result will be given if it is an inhibitor of topo I (i.e. false-positives are possible)
This is illustrated in the scheme below:
優(yōu)勢供應(yīng)產(chǎn)品：
譜析（上海）生物科技有限公司是一家致力于為實驗室分析領(lǐng)域
更多產(chǎn)品請咨詢上海熹垣生物。
This kit is designed to investigate whether a compound intercalates into the DNA double-helix, or binds in the groove, leading to unwinding of the DNA. This is a characteristic of a number of inhibitors of DNA-modifying enzymes such as the topoisomerase inhibitor m-amsacrine.
Intercalators contain planar, normally polycyclic, aromatic structures which can insert between the bases of the double-helical DNA molecule. Compounds which are able to intercalate into DNA or bind in the groove can lead to local unwinding of the DNA leading to a decrease in the twist of the DNA. If a DNA molecule is nicked and rejoined (for example by a topoisomerase) in the presence of such a compound, then the result is relaxed, underwound DNA; on removal of the enzyme and compound this converts to supercoiled DNA. The supercoiled DNA formed in these conditions is indicative of an intercalator.
This is the basis of the DNA unwinding assay. Supercoiled (or relaxed) plasmid DNA is incubated briefly with the test compound prior to relaxation by the wheat germ topo I. The enzyme and drug are then removed by extraction and the plasmid analysed by gel electrophoresis. Supercoiled topoiosmers indicate the compound is an intercalator or groove binder. If the compound is not an intercalator then the product will be relaxed plasmid.
The assay should be checked by performing a control reaction using relaxed plasmid to show that the compound is not simply acting as an inhibitor of the topo I.
The kit (product code DUKSR001) contains all you need to do the assay including:-
Supercoiled pBR322 (50 μg), relaxed pBR322 (25 μg), wheat germ topo I (250 U), Assay Buffer (1 ml) and Dilution Buffer (1 ml).
We also supply kits which contain topo I and buffers but only:-
(i) the relaxed form of pBR322 (product code DUKR002). This will show if a compound is an intercalator but not if it also inhibits the topo I (i.e. false-negatives are possible)
(ii) the supercoiled form of pBR322 (product code DUKS003). This will show if a compound is an intercalator but the same result will be given if it is an inhibitor of topo I (i.e. false-positives are possible)
This is illustrated in the scheme below:
優(yōu)勢供應(yīng)產(chǎn)品：
| Cat No. | Product |
| DUKSR001 | DNA Unwinding Assay Kit 100 assays |
| DUKR002 | DNA Unwinding Assay Kit - Relaxed only 100 assays |
| DUKS003 | DNA Unwinding Assay Kit - Supercoiled only 100 assays |
