Dithiothreitol (DTT) wani abu ne da ake amfani da shi wajen rage yawan sinadarai, wanda kuma aka sani da sabon sinadarin kore. Wani ƙaramin abu ne na kwayoyin halitta wanda ke da ƙungiyoyin mercaptan guda biyu (-SH). Saboda rage yawan sinadarai da kuma kwanciyar hankali, ana amfani da DTT sosai a gwaje-gwajen biochemistry da kwayoyin halitta.
Babban aikin DTT shine rage haɗin disulfide a cikin sunadarai da sauran ƙwayoyin halitta. Haɗin disulfide muhimmin ɓangare ne na naɗewa da kwanciyar hankali na furotin, amma a ƙarƙashin wasu yanayi na gwaji, kamar nazarin SDS-PAGE mai ragewa, haɗakar furotin da naɗewa, ya zama dole a rage haɗin disulfide zuwa ƙungiyoyi biyu na thiol don warware tsarin sararin furotin. DTT na iya amsawa tare da haɗin disulfide don rage su zuwa ƙungiyoyin mercaptan, don haka buɗe tsarin sararin furotin kuma yana sauƙaƙa bincike da sarrafawa.
Ana iya amfani da DTT don kare ayyukan enzyme da kwanciyar hankali. A wasu halayen da enzyme ke haifarwa, ana iya rage aikin enzyme ta hanyar oxidant. DTT na iya amsawa da oxidants don rage su zuwa abubuwa marasa lahani, ta haka yana kare aiki da kwanciyar hankalin enzyme.
Idan aka kwatanta da magungunan rage kiba na gargajiya kamar β-mercaptoethanol (β-ME), ana ɗaukar DTT a matsayin maganin rage kiba mafi aminci da kwanciyar hankali. Ba wai kawai yana da karko a cikin ruwan magani ba, har ma yana kiyaye halayen rage kiba a ƙarƙashin yanayin zafi mai yawa da kuma yanayin acid.
Amfani da DTT abu ne mai sauƙi. Gabaɗaya, ana narkar da DTT a cikin ma'ajiyar da ta dace sannan a ƙara shi zuwa tsarin gwaji. Ya kamata a tantance mafi kyawun yawan DTT bisa ga takamaiman gwajin, kuma gabaɗaya ana amfani da shi a cikin kewayon 0.1-1mM. Ƙananan yawan na iya rage mummunan tasirin ci gaban ƙwayoyin halitta kuma yana iya rage gubar ƙwayoyin halitta saboda yawan furotin da aka yi niyya. Babban yawan na iya haifar da nauyin metabolism na ƙwayoyin halitta mai yawa, yana shafar ci gaban ƙwayoyin halitta da ingancin bayyanar su.
Hanya mafi kyau ta tantance yawan sinadarin da aka yi amfani da shi ita ce a tantance matakin bayyanar sinadarin da aka yi amfani da shi ta hanyar gudanar da gwaje-gwajen shigar da sinadarin IPTG a wurare daban-daban. Ana iya yin gwaje-gwajen al'adu na ƙananan matakai ta amfani da nau'ikan yawan sinadarin IPTG (misali 0.1 mM, 0.5 mM, 1 mM, da sauransu) kuma ana iya kimanta tasirin bayyanar sinadarin a wurare daban-daban ta hanyar gano matakin bayyanar sinadarin da aka yi amfani da shi (misali Western blot ko gano hasken rana). Dangane da sakamakon gwaji, an zaɓi yawan sinadarin da ya fi kyawun tasirin bayyanar sinadarin a matsayin mafi kyawun yawan sinadarin.
Bugu da ƙari, za ku iya komawa ga adabin da ya dace ko ƙwarewar wasu dakunan gwaje-gwaje don fahimtar kewayon yawan IPTG da ake amfani da shi a ƙarƙashin irin waɗannan yanayin gwaji, sannan ku inganta da daidaitawa bisa ga buƙatun gwaji.
Yana da mahimmanci a lura cewa mafi kyawun yawan aiki na iya bambanta dangane da tsarin bayyanar cututtuka daban-daban, sunadaran da aka yi niyya, da yanayin gwaji, don haka ya fi kyau a inganta shi bisa ga kowane hali.
A taƙaice, DTT wani sinadari ne da ake amfani da shi wajen rage yawan sinadarin disulfide a cikin furotin da sauran ƙwayoyin halitta da kuma kare ayyukan enzymes da kwanciyar hankali. An yi amfani da shi sosai a gwaje-gwajen biochemistry da kwayoyin halitta.
Lokacin Saƙo: Satumba-28-2023