I-Hielscher Ultrasound Technology

I-Cell Lysis yamaseli we-BL21 yi-Ultrasonication

Amaseli we-BL21 awuhlobo lwe-E.coli olusetshenziswa kakhulu kumalabhorethri ocwaningo, i-biotechnology kanye nokukhiqizwa kwezimboni ngenxa yamandla abo okuveza amaprotheni asebenza kahle kakhulu. Ukuphazamiseka kwamaseli e-Ultrasonic, ukukhishwa kwe-lysis kanye namaprotheni kuyindlela ejwayelekile yokuhlukanisa nokuqoqa amaprotheni aqondisiwe avela ngaphakathi kwamangqamuzana amaseli we-BL21. I-Ultrasonication iphazamisa iseli ngokuphelele futhi ikhiphe wonke amaprotheni abambekile, okwenza i-100% yamaprotheni atholakale.

Amaseli we-BL21 Wokuvezwa Kwamaprotheni

E. coli bacteria such as BL21 cells are commonly lysed by ultrasonication in order to release expressed proteins, such as recombinant proteinsIseli le-BL21 uhlobo lwebhaktheriya olunekhono olunamakhemikhali e-E. Coli olulungele uguquko nokuvezwa kwamaprotheni asezingeni eliphezulu kusetshenziswa uhlelo lokungeniswa lwe-T7 RNA polymerase-IPTG. Amaseli we-BL21 avumela ukusebenza kahle kweprotheyini kokusebenza kwanoma isiphi isakhi esingaphansi kophromotha we-T7. Uhlobo lwe-E. coli BL21 (DE3) uhlobo lokukhiqiza amaprotheni olususelwa ku-T7 RNA oluhlanganiswe nama-vector expression asuselwa ku-T7 futhi lisetshenziswa kakhulu kumalabhorethri nasembonini ukukhiqiza amaprotheni ahlanganisayo. Ku-BL21 (DE3), ukuvezwa kofuzo olufaka iprotheni ephindaphindekayo kubhalwe yi-chromosomally encoded T7 RNA polymerase (T7 RNAP), ebhala ngokushesha okuphindwe kasishiyagalombili kune-E. Coli RNAP ejwayelekile. Lokhu kwenza uhlobo lwe-BL21 (DE3) lusebenze kahle kakhulu futhi luphenduke lube olunye lwezinhlelo zeseli ezithandwa kakhulu zamaprotheni.

Iphrothokholi ye-Ultrasonic Lysis kanye nokukhishwa kwamaprotheni kusuka kumaseli we-BL21

I-cell lysis yamaseli we-BL21 yenziwa kakhulu kusetshenziswa i-ultrasonication ngokuhlanganiswa ne-sodium lauroyl sarcosinate (eyaziwa nangokuthi i-sarkosyl) njenge-lysis buffer. Izinzuzo zokuphazamiseka kwamaseli e-ultrasonic kanye nokukhishwa kwamaprotheni kuxhomeke ekuthembekeni, ekukhiqizweni nasekusebenzeni okulula, okuphephile nokusheshayo kwama-ultrasonicators. Iphrothokholi engezansi inikeza ukuqondiswa kwesinyathelo ngesinyathelo kwe-ultrasonic BL21 cell lysis:

I-probe-type insonifier UP200St ye-lysis

Isiphazamisi seseli se-Ultrasonic UP200St nge-micro-tip S26d2 yokukhishwa kwe-lysis kanye namaprotheni

Isicelo solwazi




Qaphela kwethu Inqubomgomo yobumfihlo.


  • Ukuze kususwe amaprotheni we-chaperone, i-BL21 pellets yebhaktheriya iphinde yafakwa ku-50 ml we-ice cold Sodium Tris-EDTA (STE) buffer (equkethe i-10 mM Tris-HCL, pH 8.0, 1 mM EDTA, 150 mM NaCl engezwe nge-100 mM PMSF).
  • I-500 ul ye-lysozyme (10 mg / ml) iyangezwa futhi amaseli afakwa eqhweni imizuzu eyi-15.
  • Ngemuva kwalokho, ama-ul angu-500 we-DTT kanye no-7 ml we-sarkosyl (10% (w / v) akhiwe ku-STE buffer) ayangezwa.
  • Kubalulekile ukugcina konke ukuhlanzwa kubanda kakhulu futhi kugcine amasampula eqhweni ngaso sonke isikhathi. Zonke izinyathelo zokuhlanzwa kufanele zenziwe egumbini elibandayo uma kungenzeka.
  • VialTweeter at the ultrasonic processor UP200ST

  • Ngokukhishwa kwe-lysis ye-ultrasonic kanye nokukhishwa kwamaprotheni, amasampula enziwe nge-sonicated ku- I-VialTweeter MultiSample Ultrasonicator nge-4 x 30 sec ku-100% amplitude nge-2 min interval phakathi kwe-sonication ngayinye. Ngenye indlela, uhlobo lwe-probe homogenizer ene-micro-tip isib, UP200Ht nge-S26d2 (3 x 30 sec, 2 min. ikhefu phakathi kwemijikelezo ye-ultrasonic, 80% amplitude) ingasetshenziswa.
  • Ngezinyathelo eziqhubekayo zokuhlanzwa, amasampula kufanele agcinwe eqhweni noma ngenye indlela agcinwe ku- -80 ° C kuze kuqhutshekwe nokusebenza.

I-Ultrasonic Lysis ngaphansi kwe-Prescise Temperature Control

Ukulawulwa okuqondile nokunokwethenjelwa kokushisa kubalulekile lapho usingatha amasampula wezinto eziphilayo. Amazinga okushisa aphezulu aqala ukonakaliswa kwamaprotheni okubangelwa ukushisa ngamasampula.
Njengawo wonke amasu wokulungiselela isampula, i-sonication idala ukushisa. Kodwa-ke, izinga lokushisa lamasampuli lingalawulwa kahle lapho usebenzisa i-VialTweeter. Sikunikeza izinketho ezahlukahlukene zokuqapha nokulawula ukushisa kwamasampuli akho ngenkathi uzilungiselela nge-VialTweeter neVialPress ukuze zihlaziywe.

  1. Ukuqapha izinga lokushisa kwesampula: Iprosesa ye-ultrasonic i-UP200St, eshayela i-VialTweeter, ifakwe isoftware ehlakaniphile nenzwa yokushisa engaxhuma. Xhuma inzwa yokushisa ku-UP200St bese ufaka ichopho lenzwa yokushisa kwelinye lamashubhu esampula. Nge-digital-touch touch-display, ungasetha kumenyu ye-UP200St ibanga elithile lokushisa le-sonication yakho yesampula. I-ultrasonicator izoma ngokuzenzakalela lapho kufinyelela izinga lokushisa okuphezulu futhi imise isikhashana kuze kube yilapho izinga lokushisa lesampula lehla kunani eliphansi lokushisa okusethiwe ∆. Ngemuva kwalokho i-sonication iqala ngokuzenzekelayo futhi. Lesi sici esihlakaniphile sivimbela ukonakala okubangelwa ukushisa.
  2. Ibhlokhi ye-VialTweeter ingapholiswa ngaphambilini. Faka i-VialTweeter block (kuphela i-sonotrode ngaphandle kwe-transducer!) Efrijini noma efrijini ukupholisa ngaphambi kwebhulokhi ye-titanium kusiza ukuhlehlisa ukukhuphuka kwamazinga okushisa kusampula. Uma kunokwenzeka, isampuli uqobo lwayo ingapholiswa ngaphambilini nayo.
  3. Sebenzisa iqhwa elomile ukupholisa ngesikhathi se-sonication. Sebenzisa ithileyi elingajulile eligcwele iqhwa elomile bese ubeka i-VialTweeter eqhweni ukuze ukushisa kuphele ngokushesha.

Amakhasimende emhlabeni wonke asebenzisa i-VialTweeter ne-VialPress ngomsebenzi wabo wokulungiselela isampula nsuku zonke kumalabhorethri ezinto eziphilayo, ezamakhemikhali, ezokwelapha nezemitholampilo. Isoftware ehlakaniphile nokulawulwa kokushisa kweprosesa ye-UP200St, izinga lokushisa lilawulwa ngokuthembekile futhi nokucekelwa phansi kwesampula okubangelwa ukushisa. Ukulungiselela isampula ye-Ultrasonic ne-VialTweeter neVialPress kunikeza imiphumela ethembeke kakhulu futhi ekhiqizwayo!

Thola i-Optimal Ultrasonic Disruptor yohlelo lwakho lokusebenza lwe-Lysis

Hielscher's industrial processors of the hdT series can be comfortable and user-friendly operated via browser remote control.I-Hielscher Ultrasonics ingumkhiqizi onesipiliyoni sesikhathi eside wokuphazamisa ukusebenza kwamaseli aphezulu we-ultrasonic kanye nama-homogenizers ama-laboratories, amasistimu ebhentshi phezulu kanye nezilinganiso zezimboni. Usayizi wesiko lakho leseli lebhaktheriya, umgomo wakho wocwaningo noma wokukhiqiza kanye nevolumu yeseli elizocubungulwa ngehora noma ngosuku yizinto ezibalulekile zokuthola ukuphazamiseka kweseli kwe-ultrasonic okulungile kohlelo lwakho lokusebenza.
I-Hielscher Ultrasonics inikeza izixazululo ezahlukahlukene zokwenza i-sonication kanyekanye yamasampuli amaningi (kuze kube izitsha eziyi-10 ezine-VialTweeter) namasampuli amaningi (okungukuthi, amapuleti ama-microtiter / amapuleti e-ELISA ane-UIP400MTP), kanye ne-classic probe-type lab ultrasonicator enezinhlobo ezihlukile amazinga wamandla kusuka kuma-watts angama-50 kuye kwangama-400 kuya kuma-processor wezimboni asebenza ngokugcwele anama-16,000watts ngeyunithi ngayinye yokuphazamiseka kwamaseli ezentengiselwano kanye nokukhishwa kwamaprotheni ekukhiqizeni okukhulu. Onke ama-ultrasonicators ama-Hielscher akhelwe ukusebenza kwe-24/7/365 ngaphansi komthwalo ogcwele. Ukuqina nokuthembeka kuyizici ezisemqoka zamadivayisi wethu we-ultrasonic.
Onke ama-homogenizers edijithali e-ultrasonic afakwe isoftware ehlakaniphile, isibonisi sokuthinta esinemibala kanye neprothokholi yedatha ezenzakalelayo, eyenza idivaysi ye-ultrasonic ibe yithuluzi lomsebenzi elilula kumalab nezindawo zokukhiqiza.
Sazise, luhlobo luni lwamaseli, mthamo muni, unamaza amangaki nokuthi unaliphi ilitshe ukucubungula amasampula akho ezinto eziphilayo. Sizokuncoma ukuthi uphazamise amaseli we-ultrasonic ofanele kakhulu izidingo zakho zenqubo.

Ithebula elingezansi likunikeza inkomba yamandla wokucutshungulwa alinganiselwe amasistimu wethu we-ultrasonic kusuka kuma-homogenizers aphethwe ngesandla nama-MultiSample Ultrasonicators kuya kuma-processor we-industrial ultrasonic wezinhlelo zokuthengisa:

Iqoqwana Ivolumu flow Rate Amadivayisi ezinconyiwe
Amapuleti angama-96-kahle / ama-microtiter na I-UIP400MTP
Izitsha eziyi-10 kuya ku-0.5 kuye ku-1.5mL na I-VialTweeter ku-UP200St
0.01 kuya ku-250mL 5 kuye ku-100mL / min UP50H
0.01 kuya ku-500mL 10 kuya ku-200mL / min UP100H
10 kuya ku-2000mL 20 kuya ku-400mL / min UP200Ht, UP400St
0.1 kuya ku-20L 0.2 kuya ku-4L / min I-UIP2000hdT
10 kuya ku-100L 2 kuya ku-10L / iminithi I-UIP4000hdT
na 10 kuya ku-100L / iminithi UIP16000
na sikhulu yinhlanganisela UIP16000

Xhumana nathi! / Cela Us!

Cela ulwazi oluthe xaxa

Uyacelwa ukuthi usebenzise ifomu elingezansi ukucela ulwazi olungeziwe mayelana nezinhlelo zokusebenza ze-ultrasonic, izinhlelo zokusebenza nentengo. Sizokujabulela ukuxoxa ngenqubo yakho nawe futhi sikunikeze uhlelo lwe-ultrasonic oluhlangabezana nezidingo zakho!









Sicela uqaphele wethu Inqubomgomo yobumfihlo.


Ultrasonicator UP200Ht with microtip S26d2 for ultrasonic lysis of biological samples

I-Ultrasonicator UP200Ht nge-2mm microtip S26d2 ye-sonication yamasampuli amancane

Izincwadi / Izikhombo



Amaqiniso Okufanele Ukwazi

I-Escherichia Coli Bacteria

I-Escherichia coli wuhlobo lwebhaktheriya, okungelona elenza spore, i-Gram-negative futhi ibonakala ngesimo sayo senduku eqondile. E.coli bacteria ikhona emvelweni, ekudleni, nasemathunjini abantu nezilwane. E. coli imvamisa i-motile ngokusebenzisa i-peritrichous flagella, kepha kunezinhlobo ezingezona ezezimoto, nazo. E.coli kuthiwa yizinto eziphilayo ze-chemoorganotroph, okusho ukuthi ziyakwazi ukuphefumula nokuphefumula. Izinhlobo eziningi ze-E. Coli zinobungozi futhi zifeza imisebenzi ewusizo emzimbeni, isb ukucindezela ukukhula kwezinhlobo zamagciwane eziyingozi, ukuhlanganisa amavithamini njll.
Iseli lamagciwane i-Escherichia coli okuthiwa yi-B uhlobo oluyisigaba esikhethekile sezinhlobo ze-E. Coli, ezisetshenziswa kakhulu ocwaningweni lokuphenya izindlela ezinjengokuzwela kwe-bacteriophage noma amasistimu wokuguqula ukukhawulelwa. Ngaphezu kwalokho, amabhaktheriya e-E. Coli athathwa njengobungcweti obunokwethenjelwa bokuveza amaprotheni ku-biotechnology nakuma laboratories esayensi yokuphila. Isibonelo, u-E. Coli asetshenziselwa ukuhlanganisa izinto ezifana namaprotheni nama-oligosaccharides esikalini sezimboni. Ngenxa yezici ezithile ezifana nokushoda kweprotease, ukukhiqizwa kwe-acetate ephansi ezingeni eliphakeme le-glucose, nokutholakala okuthuthukisiwe, amangqamuzana e-E coli B ngawona asetshenziswa kakhulu amangqamuzana okusingathwa ekukhiqizeni amaprotheni enzelwe izakhi zofuzo.

Amaphrotheni Aphindayo

Amaprotheni ahlanganisayo (rProt) athola ukubaluleka okukhulu emagatsheni amaningi, kufaka phakathi ukukhiqizwa kwamakhemikhali, okwenziwa ngamakhemikhali, izimonyo, imithi yabantu neyezilwane, ezolimo, ukudla kanye nezimboni zokwelapha imfucuza.
Ukukhiqizwa kwamaprotheni ahlanganisayo kudinga ukusetshenziswa kohlelo lokuveza. Njengokuveza amasistimu amaseli wokukhiqizwa kwe-DNA ephindaphindayo, amaseli we-prokaryotic ne-eukaryotic angasetshenziswa. Ngenkathi iseli le-bacterial lisetshenziswa kakhulu ekuvezeni amaprotheni ngenxa yezinto ezinjengezindleko eziphansi, ukusabalalisa kalula nezimo ezilula zemidiya, ezinye izindlela ezisunguliwe ziyimammalia, imvubelo, ulwelwe, izinambuzane namaseli angenamaseli. Uhlobo lwamaprotheni, umsebenzi osebenzayo, kanye nesivuno esidingekayo samaprotheni aveziwe sinomthelela ekukhetheni kohlelo lweseli olusetshenziselwa ukubonakaliswa kwamaprotheni.
Ukuze kuvezwe amaprotheni ahlanganisayo, iseli elithile kufanele lithunyelwe nge-vector ye-DNA equkethe isifanekiso se-DNA ephindaphindekayo. Amaseli adluliselwe ngesifanekiso ayalungiswa. Njengomphumela womshini weselula, amaseli abhala futhi ahumushe iphrotheni lentshisekelo, ngaleyo ndlela akhiqize iphrotheni elihlosiwe.
Njengoba amaprotheni aveziwe abhajwe kumatrix weselula, iseli kufanele lifakwe intambo (liphazanyiswe futhi liphuke) ukukhipha amaprotheni. Esikhathini esilandelayo sokuhlanzwa, amaprotheni ahlukaniswe futhi ahlanzwe.
Iprotheyini yokuqala yokwakha kabusha esetshenziswe ekwelashweni kwakuyi-insulini yomuntu ephindayo ngo-1982. Namuhla, kunezinhlobo ezingaphezu kuka-170 zamaphrotheni ahlanganiswa kabusha emhlabeni wonke ezenzelwe ukwelashwa. Amaprotheni asetshenziswayo ajwayelekile asetshenziswa kwezokwelapha ngokwesibonelo ama-hormone aphindaphindayo, ama-interferon, ama-interleukin, izici zokukhula, izinto zesimila necrosis, izinto eziqinisa igazi, izidakamizwa ze-thrombolytic, nama-enzyme ekwelapheni izifo ezinkulu njengesifo sikashukela, ubufishithi, i-myocardial infarction, isifo sokucinana kwenhliziyo, i-cerebral apoplexy, i-multiple sclerosis, i-neutropenia, i-thrombocytopenia, i-anemia, i-hepatitis, i-rheumatoid arthritis, i-asthma, isifo sikaCrohn kanye nemithi yokwelapha yomdlavuza. (cf.Phuc V. Pham, ku-Omics Technologies naku-Bio-Engineering, 2018)


Hielscher Ultrasonics supplies high-performance ultrasonic homogenizers from lab to industrial size.

Ukusebenza okuphezulu kwe-ultrasonics! Uhla lomkhiqizo kaHielscher lumboza isibonisi esigcwele kusuka ku-compact lab ultrasonicator ngaphezulu kwamayunithi ebhentshini-phezulu kuya ezinhlelweni ezigcwele zezimboni ze-ultrasonic.