Thiazolyl Blue CAS 298-93-1

Shaanxi BLOOM Tech Co., Ltd. yog ib tug ntawm cov uas paub txog manufacturers thiab lwm tus neeg ntawm thiazolyl xiav cas 298-93-1 nyob rau hauv Suav teb. Txais tos rau cov lag luam wholesale bulk zoo thiazolyl xiav cas 298-93-1 muag ntawm no los ntawm peb lub hoobkas. Kev pabcuam zoo thiab tus nqi tsim nyog muaj.

Thiazolyl xiav, zoo dua lub npe hu ua MTT (3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide), harbors ib tug fascinating duality-nws yog ob qho tib si tseem ceeb hauv cell biology thiab ib tug chameleonic tshuaj oddity. Cov hmoov daj crystalline no dhau los ua qhov pib metamorphosis hauv cov hlwb nyob: mitochondrial reductases cleave nws lub nplhaib tetrazolium, hloov nws mus rau hauv insoluble liab doog formazan precipitate-ib qho tshuaj tiv thaiv kom pom kev tawm tsam tias nws tau dhau los ua tus qauv kub rau kev ntsuas ntawm tes. Tsis tau dhau ntawm lub rooj zaum hauv chaw sim, MTT nthuav tawm ntau tus cwj pwm esoteric; nws cov qauv crystalline tsim tshwj xeeb "herringbone" qauv nyob rau hauv microscopy, thaum nws cov khoom redox tso cai rau nws koom nrog cov tshuaj tiv thaiv kab mob sib txawv thaum raug rau cov wavelengths ntawm lub teeb. Curiously, lub compound nthuav tawm qhov tsis tshua muaj tshwm sim hu ua "metachromasia" - tshwm sim xim sib txawv nyob ntawm qhov concentration thiab molecular ib puag ncig-ib qho cuab yeej siv los ntawm textile chemists los tsim "ntse" dyes. Cov kev tshawb fawb tsis ntev los no tau nthuav tawm nws lub luag haujlwm tsis txaus ntseeg raws li qhov tsis muaj zog inhibitor ntawm qee cov kab mob ua pa enzymes, hinting tias qhov kev ntsuam xyuas ubiquitous reagent no tau cuam tshuam rau cov txheej txheem metabolic uas cov kws tshawb fawb siv nws los kawm. Thaum yaj nyob rau hauv cov kuab tshuaj organic, MTT cov kev daws teeb meem pom qhov txawv txav fluorescence quenching nyhuv uas muaj puzzled lub cev chemists, qhia complex intermolecular kev sib tshuam tseem tos tag nrho cov lus piav qhia.

Thiazolyl Blue, Lub npe tshuaj yog 3- (4,5-dimethyl-2-thiazole) -2,5-diphenyltetrazolium bromide, abbreviated li MTT, uas yog ib qho tseem ceeb organic compound. Txij li thaum Mosman thawj zaug qhia txog kev siv MTT staining los txiav txim siab ntawm cell loj hlob thiab ciaj sia nyob rau hauv 1983, MTT tau siv dav hauv cov haujlwm xws li kev tshawb fawb txog kev ua neej, kev tshawb fawb kho mob, thiab kev tsim tshuaj. Cov hauv qab no yog cov lus piav qhia ntxaws txog nws lub hom phiaj:

Daim ntawv thov hauv Cell Biology Research

 

MTT yog ib qho ntawm cov txheej txheem classic rau kev txiav txim siab ntawm cell proliferation. Thaum lub sij hawm cell kab lis kev cai, dehydrogenases xws li succinate dehydrogenase nyob rau hauv cell mitochondria yuav txo tau exogenous MTT rau insoluble xiav ntshav liab crystalline formazan, uas deposits nyob rau hauv lub cell. Cov hlwb tuag tsis muaj peev xwm txo qhov no thiab yog li tsis tsim cov formazan. Los ntawm dissolving formazan nyob rau hauv hlwb thiab ntsuas nws lub teeb haum tus nqi ntawm ib lub wavelengths (xws li 490nm los yog 562nm) siv ib tug enzyme -linked immunosorbent assay (ELISA) nyeem ntawv, tus naj npawb ntawm cov nyob rau hauv lub hlwb tuaj yeem cuam tshuam ncaj qha. Nyob rau hauv ib qho ntawm cov xov tooj ntawm tes, tus nqi ntawm MTT crystals tsim yog proportional rau cov xov tooj ntawm tes, yog li cell proliferation yuav txiav txim raws li qhov ntsuas absorbance tus nqi (OD tus nqi).

Daim ntawv thov hauv Cell Biology Research

 

MTT kuj tseem tuaj yeem siv los txiav txim siab txog cov tshuaj toxicity los yog lwm txoj kev kho mob (xws li hluav taws xob irradiation) ntawm kab lis kev cai hauv vitro. Los ntawm kev sib piv cov txiaj ntsig OD ntawm pab pawg kho mob thiab pab pawg tswj hwm, cov txiaj ntsig ntawm cov tshuaj lossis cov txheej txheem kho mob ntawm tes tuaj yeem soj ntsuam. Qhov loj dua tus nqi OD, lub zog ua haujlwm ntawm tes; Ntawm qhov tsis sib xws, nws qhia tau hais tias qhov tsis muaj zog ntawm cov haujlwm ntawm tes lossis ntau dua ntawm cov cell toxicity. Txoj kev no muaj kev siv dav hauv kev tshuaj xyuas tshuaj, kev tshawb fawb toxicology, thiab lwm yam. Txawm hais tias MTT feem ntau yog siv los ntsuas kev loj hlob ntawm tes thiab cytotoxicity, qee zaum, nws kuj tuaj yeem siv los pab hauv kev tshawb fawb ntawm cell apoptosis. Thaum lub sij hawm tus txheej txheem ntawm cell apoptosis, cov enzyme kev ua nyob rau hauv lub cell hloov, uas tej zaum yuav cuam tshuam lub peev xwm ntawm MTT. Los ntawm kev sib piv qhov sib txawv ntawm OD qhov tseem ceeb ntawm cov hlwb apoptotic thiab cov hlwb ib txwm muaj, qhov tshwm sim ntawm cell apoptosis tuaj yeem txiav txim siab ua ntej. Txawm li cas los xij, nws yuav tsum tau muab sau tseg tias MTT tsis yog ib txoj hauv kev tshwj xeeb rau kev kuaj ntshav apoptosis, thiab nws cov txiaj ntsig yuav tsum tau muab tshuaj ntsuam xyuas ua ke nrog rau lwm txoj hauv kev tshawb nrhiav apoptosis.

Daim ntawv thov hauv kev tsim tshuaj

 

Nyob rau hauv tus txheej txheem ntawm kev txhim kho tshuaj, MTT assay yog ib tug nquag siv siab -throughput tshuaj ntsuam xyuas txoj kev. Los ntawm kev sib koom ua ke cov tshuaj ntsuam xyuas nrog cov hlwb thiab siv MTT qhov kev ntsuam xyuas los ntsuas kev hloov pauv hauv kev ua haujlwm ntawm tes, cov tshuaj muaj peev xwm ntawm cov tshuaj molecules tuaj yeem kuaj tau sai. Txoj kev no muaj qhov zoo ntawm kev ua haujlwm yooj yim, txhim khu kev ntseeg tau, thiab kev ua haujlwm siab, thiab muaj qhov tseem ceeb ntawm daim ntawv thov tus nqi nyob rau theem pib ntawm kev tsim tshuaj. Kev ntsuam xyuas MTT kuj tuaj yeem siv los kawm txog kev ua haujlwm ntawm cov tshuaj. Los ntawm kev sib piv cov kev hloov pauv ntawm kev ua haujlwm ntawm tes nyob rau hauv cov kev kho mob sib txawv, qhov cuam tshuam ntawm cov tshuaj ntawm cov txheej txheem xws li cell proliferation thiab apoptosis tuaj yeem txiav txim siab ua ntej. Los ntawm kev sib koom ua ke lwm cov txheej txheem biology molecular xws li Western blot thiab ntws cytometry, lub hom phiaj thiab cov cim qhia ntawm cov tshuaj tuaj yeem raug nthuav tawm ntxiv. Hauv kev soj ntsuam kev nyab xeeb ntawm tshuaj, MTT kev soj ntsuam tuaj yeem siv los ntsuas cov tshuaj lom ntawm cov tshuaj ntawm cov hlwb. Los ntawm kev ntsuas OD tus nqi hloov pauv ntawm cov hlwb tom qab kev kho tshuaj, nws tuaj yeem txiav txim siab seb cov tshuaj puas muaj cytotoxicity thiab lub zog ntawm toxicity. Qhov no yog qhov tseem ceeb heev rau kev qhia kev tshawb fawb preclinical thiab kev siv tshuaj kho mob.

Kev thov ntawm MTT hauv lwm qhov chaw

 

Hauv kev tshawb fawb txog kev tiv thaiv kab mob, MTT kev soj ntsuam tuaj yeem siv los txiav txim siab qhov muaj peev xwm loj hlob ntawm lymphocytes. Los ntawm kev sib koom ua ke lymphocytes nrog antigens lossis immunostimulants thiab ntsuas kev hloov pauv hauv kev ua haujlwm ntawm tes siv MTT kev soj ntsuam, lub cev tiv thaiv kab mob muaj peev xwm ntawm lymphocytes tuaj yeem soj ntsuam. Txoj kev no muaj kev siv dav hauv kev tsim tshuaj tiv thaiv, immunotherapy, thiab lwm yam. Hauv kev tshawb fawb oncology, MTT kev soj ntsuam tuaj yeem siv los txiav txim siab qhov kev loj hlob thiab kev siv tshuaj rhiab heev ntawm cov qog hlwb. Los ntawm kev sib piv cov kev hloov pauv hauv OD qhov tseem ceeb ntawm cov qog hlwb nyob rau hauv cov kev kho mob sib txawv, cov tshuaj molecules nrog inhibitory cuam tshuam ntawm cov qog hlwb tuaj yeem raug tshuaj xyuas thiab lawv cov tshuaj tiv thaiv- qog tuaj yeem soj ntsuam. Qhov no yog qhov tseem ceeb heev rau kev taw qhia kev kho mob ntawm cov qog. Hauv kev tshawb fawb txog toxicology ib puag ncig, MTT kev soj ntsuam tuaj yeem siv los ntsuas cov tshuaj lom ntawm ib puag ncig cov pa phem ntawm cov hlwb. Los ntawm kev sib koom ua ke cov hlwb nrog ntau qhov sib txawv ntawm cov pa phem thiab ntsuas kev hloov pauv hauv kev ua haujlwm ntawm tes siv MTT kev soj ntsuam, cov tshuaj lom thiab cov txheej txheem ntawm kev ua pa phem tuaj yeem txiav txim siab. Qhov no yog qhov tseem ceeb heev rau kev soj ntsuam ecological txaus ntshai ntawm ib puag ncig cov pa phem thiab tsim cov cai tiv thaiv ib puag ncig. 

Kev sim cov kauj ruam ntawmthiazolyl xiavtxoj kev:

1: Trypsin digests logarithmic theem hlwb, centrifuges thiab sau lawv tom qab txiav tawm kom tshem tawm ntawm tes, thiab kho nws cov concentration rau 5-10 los ntawm cell count × 104 / ml. Rau cov pib tshiab, nws yog ib qho tsim nyog yuav tsum ua kom cov hlwb ncav cuag 5-10 × 104 / ml feem ntau tsis paub yuav pib qhov twg. Kuv nyob ntawm no los muab ib txoj hauv kev yooj yim rau koj los txiav txim seb tus lej ntawm cov cell.

Siv 25cm2 feem ntau siv rau kab lis kev cai ntawm tes ua piv txwv:

1) Cell ceev yog li ntawm 80% ~ 90% (daim duab hauv qab no qhia cov kwv yees lub xeev ntawm cov cell ntawm 80% ~ 90% ntom). Tom qab kev zom thiab centrifugation, tshem tawm cov supernatant, thiab ntxiv 3ml nruab nrab rau nws.

2) Siv 15ml sterile centrifuge raj tshiab (rau cov kauj ruam tom ntej ntawm inoculation ntawm 96-zoo phaj) thiab sau nws nrog li 9ml nruab nrab

3) Siv 1ml los ntawm 3ml cell suspension npaj rau thawj kauj ruam, ntxiv rau hauv lub raj sab sauv, thiab suav cov hlwb tom qab sib tov. Nyob rau lub sijhawm no, nws feem ntau yog 5-10 lossis tsawg dua × 104 / ml, uas yog sib npaug rau 5-10 hlwb ntawm ib lub xovtooj ntawm 4 lub hlwb ntawm lub xov tooj ntawm tes suav nrog (saib daim duab hauv qab no); Yog tias qhov concentration tsis txaus, qhov kev ncua ntawm tes yuav tsum tau ntxiv raws li qhov suav tau, thiab txhua qhov poob yuav raug suav tias yog 50ul.

4) Tus naj npawb ntawm cov hlwb yuav tsum raug kho raws li qhov sib txawv ntawm qhov kev sim. Piv txwv li, nyob rau hauv kev sim loj hlob ntawm tes, 2000 hlwb ntawm ib qhov dej yuav siv tau (sib npaug rau cell suspension ceev ntawm 2 × 104 / ml), 5000 mus rau 10000 hlwb ib qhov dej (sib npaug rau cell suspension ceev ntawm 5 × 104 / ml). Tsis tas li ntawd, tus naj npawb ntawm cov cell yuav tsum tau txiav txim siab raws li lawv tus kheej cov yam ntxwv, xws li kev loj hlob ceev. Piv txwv li, qhov ceev ntawm cov cell loj hlob sai yuav me ntsis me ntsis.

2. Tom qab npaj lub cell suspension, sib tov maj mam thiab ntxiv 100ul rau hauv txhua lub qhov, kom qhov ceev ntawm cov cell yuav tsum tau kuaj yog 5000-10000 / qhov (lub qhov ntug yog ntim nrog sterile PBS).

L Nco tseg: Vim tias cov hlwb yuav nyob twj ywm tom qab sib tov, nws yog ib qho tsim nyog yuav tsum sib tov ntau zaus thaum lub sij hawm inoculation, xws li sib tov ib zaug txhua rau lub qhov, kom ntseeg tau tias lub cell ceev ntawm inoculation yog raws nraim tib yam ntawm lub qhov, uas yog ib qho tseem ceeb rau cov kev tshwm sim ntawm MTT.

3. Muab cov inoculated cell kab lis kev cai phaj rau hauv incubator rau cultivation kom txog rau thaum lub cell monolayer yog them nrog rau hauv qab ntawm lub qhov (96-qhov ca phaj), thiab ntxiv cov tshuaj nrog concentration gradient. Raws li txoj cai, cov tshuaj tuaj yeem muab ntxiv tom qab lub xov tooj ntawm tes txuas rau ntawm phab ntsa, lossis ob teev, lossis ib nrab hnub, tab sis peb feem ntau ntxiv cov tshuaj nyob rau yav tav su ntawm hnub dhau los, thiab tag kis sawv ntxov. Feem ntau, 5-7 gradients, 100ul ib lub qhov, thiab 3-5 ntau qhov yog teem. Nws raug pom zoo kom teeb tsa 6, txwv tsis pub nws nyuaj rau xav txog qhov xwm txheej tiag tiag. Cov duab hauv qab no tuaj yeem siv los ua cov kauj ruam siv rau 96-qhov phaj.

L Rau kev siv tshuaj, qee tus neeg ncaj qha ntxiv cov tshuaj rau hauv 96-zoo daim hlau los ntawm ntau qhov sib txawv los ua kom muaj kev sib haum xeeb. Tab sis kuv xav tias peb yuav tsum sim sib xyaw cov tshuaj sib txawv hauv EP raj, tom qab ntawd tshem tawm cov kab lis kev cai supernatant los ntawm 96-zoo phaj (uas tuaj yeem nqus tau nrog rab phom tso tawm) thiab ntxiv 100ul ntawm kab lis kev cai nruab nrab uas muaj cov tshuaj sib txawv, uas tuaj yeem ua kom qhov tseeb ntawm cov tshuaj concentration. Tsis tas li ntawd, nws yuav tsum tau muab sau tseg tias yog siv txoj kev no, tsis txhob nqus tag nrho 96-zoo phaj kab lis kev cai tov ua ntej muab tshuaj. Koj yuav tsum ntxiv cov qauv tam sim ntawd tom qab nqus ib feem ntawm nws kom tsis txhob muaj cell tuag vim ziab ntawm 96-zoo phaj.

Incubate nrog 4.5% CO2 ntawm 37℃rau 16-48 teev, thiab saib xyuas cov nyhuv ntawm cov tshuaj nyob rau hauv lub inverted microscope. Cov duab hauv qab no qhia txog cov tshuaj gradient ntawm cytotoxicity.

4. Ntxiv 10ul MTT tov (5mg/ml, piv txwv li . 0.5% MTT) rau txhua qhov dej thiab txuas mus rau kab lis kev cai rau 4h. Yog tias cov tshuaj tuaj yeem hnov ​​​​mob nrogthiazolyl xiavcov kab lis kev cai tuaj yeem muab pov tseg tom qab centrifugation, thiab kab lis kev cai daws teeb meem uas muaj MTT tuaj yeem muab ntxiv tom qab ntxuav nrog PBS rau 2-3 zaug.

5. Tso tseg cov kab lis kev cai thiab npaj rau kev tawg thiab crystallization.

Thiazolyl Blue, uas nws lub npe Suav yog Methyl Blue, yog ib qho tseem ceeb organic compound uas yog dav siv hauv tshuaj, biology thiab chemistry. Hauv cov tshuaj, nws feem ntau yog siv los kho cov tshuaj lom nitrite, tshuaj lom cyanide, thiab lwm yam, thiab kuj tseem siv rau hauv kev kuaj mob thiab kev kho mob ntawm qee yam kab mob; Hauv kev tshawb fawb lom neeg, nws tuaj yeem siv los ua tus neeg sawv cev staining rau kev sau cov hlwb lossis cov ntaub so ntswg. Txawm li cas los xij, zoo li ntau yam tshuaj, methylene xiav kuj tseem muaj cov kev mob tshwm sim thaum siv:

Digestive system phiv

Kev xeev siab thiab ntuav yog ib qho ntawm cov kab mob plab zom mov tshwm sim ntawm methylene xiav. Thaum methylene xiav nkag mus rau hauv lub cev, nws yuav ua rau lub plab hnyuv mucosa, ua rau xeev siab thiab ntuav thaum mob hnyav. Cov tsos mob no tau tshaj tawm thaum qhov kev txhaj tshuaj ntawm cov hlab ntsha nrawm dhau los yog qhov ntau npaum li cas. Piv txwv li, hauv qee qhov chaw kho mob, cov neeg mob yuav hnov ​​​​qhov xeev siab thiab ntuav sai sai tom qab txhaj tshuaj methylene xiav, uas cuam tshuam rau lawv txoj kev nplij siab thiab kev kho mob. Qee tus neeg mob yuav hnov ​​​​mob plab tom qab siv cov tshuaj methylene xiav, thiab qhov mob ntawm qhov mob yuav txawv nyob ntawm tus neeg sib txawv, xws li qhov tsis xis nyob mus rau qhov mob hnyav dua.

Nyob rau tib lub sijhawm, nws kuj tseem tuaj yeem ua rau raws plab, uas tej zaum yuav yog vim muaj kev cuam tshuam ntawm methylene xiav ntawm plab hnyuv peristalsis thiab digestive functions, ua rau lub plab zom mov txawv txav thiab kev hloov pauv hauv plab hnyuv secretion.Ntau zaus los yog nquag tshwm sim ntawm qhov mob plab thiab raws plab, tab sis tus neeg mob plab tsis tuaj yeem cuam tshuam rau kev noj qab haus huv xwb. rau lwm yam teeb meem.

Urinary system phiv

Tom qab siv methylene xiav, cov zis xim yuav tig xiav, uas yog tshwm sim los ntawm kev tso tawm ntawm methylene xiav thiab nws cov metabolites rau hauv cov zis los ntawm ob lub raum. Feem ntau, qhov kev hloov ntawm cov zis xim no yog ib ntus thiab yuav tsis muaj kev cuam tshuam loj rau kev noj qab haus huv ntawm lub cev. Tab sis qee qhov tshwj xeeb, xws li cov neeg mob uas muaj kab mob hauv lub cev, kev hloov pauv ntawm cov zis xim tuaj yeem cuam tshuam nrog kev kuaj mob ntawm tus kab mob, thiab cov kws kho mob yuav tsum tau ua qhov kev ntsuam xyuas dav dav raws li lwm yam kev ntsuas ntsuas. Thaum tso zis, qee cov neeg mob yuav hnov ​​​​lub suab nrov ntawm qhov qhib urethral.

 

Qhov no tej zaum yuav yog vim cov nyhuv stimulating ntawm methylene xiav ntawm lub urethral mucosa, tshwj xeeb tshaj yog thaum cov zis concentration siab los yog lub urethral mucosa yog rubbed thaum tso zis, stinging hnov ​​​​ntxhiab heev. Cov tsos mob no tuaj yeem ua rau qee qhov tsis xis nyob rau cov neeg mob thiab cuam tshuam rau lawv lub neej txhua hnub.

Neurological phiv

kiv taub hau thiab mob taub hau yog ib qho kev mob tshwm sim hauv paj hlwb ntawm methylene xiav. Tom qab siv methylene xiav, cov neeg mob yuav hnov ​​​​zoo li kiv taub hau thiab tsis xis nyob hauv lub taub hau, lossis muaj kev tsis xis nyob xws li o thiab mob taub hau. Cov tsos mob no yuav cuam tshuam rau tus neeg mob txoj kev xav thiab kev muaj peev xwm, thiab txo qis kev ua haujlwm thiab kev ua haujlwm ntawm lub neej. Qhov tshwm sim ntawm kiv taub hau thiab mob taub hau tuaj yeem cuam tshuam txog cov teebmeem ntawm methylene xiav ntawm lub hlwb cov hlab ntsha thiab cov hlab ntsha, thiab cov txheej txheem tshwj xeeb tseem xav tau kev tshawb fawb ntxiv. Muaj tsawg zaus, methylene xiav yuav ua rau muaj kev puas siab puas ntsws xws li qaug zog, coma, thiab lwm yam.

Qhov no feem ntau yog vim kev siv ntau dhau ntawm methylene xiav los yog cov neeg mob ua rau cov tshuaj ntau dhau, ua rau muaj kev cuam tshuam loj heev ntawm lub hauv paus paj hlwb. Kev puas siab puas ntsws yog ib qho kev mob tshwm sim loj uas yuav ua rau tus neeg mob txoj sia muaj kev nyab xeeb thiab xav tau kev kho mob sai sai.

1.Dab tsi yog kev siv thiazolyl xiav tetrazolium bromide?

Thiazolyl Blue Tetrazolium Blue (MTT) tuaj yeem siv los ntsuas kev loj hlob ntawm tes. MTT tsim cov tshuaj daj daj uas hloov mus rau xiav tsaus, dej -insoluble MTT formazan los ntawm mitochondrial dehydrogenases ntawm cov cell nyob.

2.Puas Thiazolyl Blue muaj kev ruaj ntseg rau khaws cia?

Qhov kub thiab txias: kaw cov hmoov thiab muab tso rau hauv lub tub yees ntawm -20℃rau kev khaws cia khov, uas tuaj yeem ua kom ntev lub neej txee (feem ntau yog 1-2 xyoos); rau lub sijhawm luv luv (hauv 1 lub hlis), nws tuaj yeem khaws cia ib ntus hauv qab 4℃tub yees.

Lub teeb- pov thawj thiab noo noo- Cov ntaub ntawv pov thawj: MTT hmoov yog lub teeb - rhiab heev thiab yuav tsum muab khaws cia rau hauv cov fwj xim av reagent lossis lub raj mis pob tshab qhwv nrog cov ntawv ci. Lub caij no, cov neeg ua kom qhuav (xws li, silica gel) yuav tsum tau ntxiv los tiv thaiv cov hmoov los ntawm absorbing noo noo thiab caking.

3.Puas yuav Thiazolyl Blue xim hloov?

Thaum cov kev daws teeb meem pom cov xim xiav xim daj los yog tsaus xim, nws qhia tau tias nws tau oxidized thiab tsis tuaj yeem siv txuas ntxiv mus.
Tsis txhob muab cov tshuaj tso rau ntawm chav tsev kub lossis hauv qhov chaw teeb pom kev zoo, vim nws yuav ua tsis tau sai.

Cim npe nrov: thiazolyl blue cas 298-93-1, lwm tus neeg, manufacturers, hoobkas, lag luam wholesale, yuav, nqe, tej, kev muag khoom