dynamic BIOSENSORS Normalisation Solution Software

Nkọwapụta

• Aha ngwaahịa: helicity
• Ụdị: Ngwọta Normalization (ihe na-acha uhie uhie)
• Maka: Nyocha RT-IC na ọwa uhie
• Onye nrụpụta: Dynamic Biosensors GmbH & Inc.
• Nọmba ịtụ ahịa: NOR-Ra
• Maka Nnyocha Jiri naanị
• Ndụ nchekwa: Oke, lelee ụbọchị njedebe na labelụ

Ihe ndị na-emetụta ike mkpali:

1. Fluorophore itinye uche na ngwọta nyocha
2. Mgbama njide tụrụ anya
3. Ụdị mgbawa

Maka ntụzịaka akọwapụtara nke ọma na ike mkpali na nhazi nhazi nhazi, rụtụ aka na Tebụl 2 enyere na ntuziaka onye ọrụ. Rịba ama na enwere ike ịchọ nkwalite ụfọdụ dabere na sistemu onye ọ bụla.

Ajụjụ

• Ajụjụ: Kedu ka m ga-esi chekwaa helicity?
  • A: Chekwaa heliXcyto dịka ozi nchekwa enyere na ntuziaka onye ọrụ. Gbaa mbọ hụ na ịlele ụbọchị njedebe na labelụ ma jiri ya tupu ngafe.
• Ajụjụ: Enwere ike iji helicity maka ebumnuche ụlọ ọgwụ?
  • A: Mba, heliXcyto bụ maka nyocha naanị na ekwesighi iji ya maka nyocha ụlọ ọgwụ ma ọ bụ ọgwụgwọ.

Atụmatụ igodo

• Maka nhazi nke akara fluorescent na Spot 1 na Spot 2 nke mgbawa heliXcyto.
• Na-enyere aka ntutụ aka ziri ezi nke mgbama fluorescent na-acha uhie uhie n'oge nha RT-IC
• Dakọtara na ibe helicity niile
• Ngwọta Normalisation (ihe na-acha uhie uhie) nwere ihe na-acha uhie uhie hydrophilic nke nwere otu ụgwọ net dị mma.

Nkọwa ngwaahịa

• Nọmba ịtụ ahịa: NOR-Ra

Tebụl 1. Ọdịnaya na Ozi Nchekwa

Ihe onwunwe	Okpu	Ihe omume uche	Ọnụ ego	Nchekwa
Normalization ngwọta-Ra	oroma	10 µM	6 x 100 µL	-20 Celsius

• Maka iji naanị nyocha.
• Ngwaahịa a nwere ndụ nchekwa nwere oke, biko hụ ụbọchị ngwụcha na label.

Nkwadebe

• Jiri ihe ngwọta normalization dye red a maka nha RT-IC na ọwa uhie (analyte label-based).
• Gwakọta 10 μM normalization ngwaahịa ngwọta na ntinye uche na-arụ ọrụ na ihe nchekwa na-agba ọsọ.
• Nleba anya nke ngwọta normalization kwesịrị ịdị nhata kwekọrọ na ntinye fluorophore na mkpokọta nyocha kachasị elu ka a tụrụ.

Enwere ike gbakọọ nke a site na iji nha nha:

• : Ntinye uche nke ngwọta normalization na agba a chọrọ
• : Ntụkwasị nke agba na ngwọta nyocha akpọrọ
• : Ntụle nyocha kachasị elu nke kwesịrị ịtụ
• : ogo nke labeling (oke nke agba ka nyocha)

Enwere ike ịchekwa ihe ngwọta a gwakọtara na 2-8 Celsius C ruo ụbọchị 7.

Ngwa ndetu

Na nha RT-IC, mgbaàmà fluorescent nke ngwọta normalization kwesịrị ịdị n'ụdị yiri nke ahụ dị ka mgbama kachasị elu na-abịa site na bound analyte (raw data). Mgbama fluorescent zuru oke na-adabere na nhazi nhazi nhazi nhazi yana ike mkpali etinyere na nha. A ga-ahọrọ ike mkpali dabere na paramita ndị a:

• a. Ntinye uche nke fluorophore na ngwọta nyocha:
  • Ntinye uche nke fluorophore dabere na nleba anya nyocha ejiri na nha yana ogo akara nke nyocha. Maka nnukwu DOL na mkpokọta nyocha dị elu, enwere ike ịchọ iweda ike mkpali.
• b. Mgbama njide tụrụ anya:
  • Ebumnobi egosipụtara nke ukwuu na cell nwere ike jikọta ụmụ irighiri ihe nke nyocha akpọrọ. N'ihe gbasara ebumnobi nke ukwuu, enwere ike ịtụ anya mgbama njikọ siri ike. Iji zere mmechi mkpuchi, enwere ike ịtụle iweda ike mkpali.
• c. Ụdị mgbawa:
  • Ụdị mgbawa dị iche iche nwere ọdịdị fluorescent dị iche iche. Ka ọnyà ndị ahụ buru ibu na ọnyà ndị ọzọ na mgbawa, ka mgbaàmà ndabere dị elu. Ya mere, ibe L5 nwere ike ịchọ ike mkpali dị ala karịa etinyere na ibe M5.

Maka mmalite nke ike mkpali na ụkpụrụ. Ngwọta ngwọta nke a ga-eji mee ihe na nnwale RT-IC, biko rụtụ aka na Tebụl 2. Isiokwu 2. Mmekọrịta nke ịta ahụhụ fluorophore, nhazi nhazi nhazi nhazi, na ike mkpali kwesịrị ekwesị maka mgbawa heliXcyto M5.

Analyte dye conc. = analyte cons x DOL	Ike mkpali	Ngwọta nhazi nhazi nke uche	Dilution Normalisation ngwọta
25 nM	0.5	25 nM	1:400
50 nM	0.3	50 nM	1:200
100 nM	0.2	100 nM	1:100
300 nM	0.1	300 nM	1:33
500 nM	0.08	500 nM	1:20
1 µM	0.05	1 µM	1:10
2.5 µM	0.02	2.5 µM	1:4

Mara: Tebụl a bụ maka ntụzịaka gị. Otú ọ dị, mgbaàmà ikpeazụ e dekọrọ na helicity na-adabere n'ọtụtụ ihe. Ya mere, a ga-achọ ụfọdụ njikarịcha maka sistemụ ọ bụla.

Kpọtụrụ

• Dynamic Biosensors GmbH
• Perchtinger Str. 8/10
• 81379 Munich
• Germany
• Ọnọdụ nke Dynamic Biosensors, Inc.
• 300 Trade Center, Suite 1400
• Woburn, MA 01801
• USA
• Ozi ịtụ order@dynamic-biosensors.com.
• Nkwado ndị teknuzu support@dynamic-biosensors.com.
• www.dynamic-biosensors.com.
• A na-arụ ọrụ na ibe na-arụ ọrụ na Germany.
• ©2024 Dynamic Biosensors GmbH
• Dynamic Biosensors, Inc. Ikike niile echekwabara.
• NOR-Ra v1.0
• www.dynamic-biosensors.com.
• Dynamic Biosensors GmbH & Inc.
• NOR-Ra v1.0

Akwụkwọ / akụrụngwa

dynamic BIOSENSORS Normalisation Solution Software [pdf] Ntuziaka onye ọrụ NOR-Ra, Ngwọta Ngwọta Nhazi, Ngwa Ngwọta, Ngwanrọ

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