{"id":248,"date":"2018-09-09T01:45:20","date_gmt":"2018-09-09T01:45:20","guid":{"rendered":"http:\/\/orobanche.net\/?p=248"},"modified":"2022-02-08T15:30:05","modified_gmt":"2022-02-08T15:30:05","slug":"enzim-aktivite-olcumleri-icin-ezme-tamponu-hazirlanmasi","status":"publish","type":"post","link":"https:\/\/orobanche.net\/?p=248&lang=tr","title":{"rendered":"Enzim Aktivite \u00d6l\u00e7\u00fcmleri \u0130\u00e7in Ezme Tamponu Haz\u0131rlanmas\u0131"},"content":{"rendered":"

Ba\u015flamadan \u00f6nce her zaman dedi\u011fim gibi literat\u00fcre iyice bir bak\u0131n. Baz\u0131 durumlarda farkl\u0131 bitkiler i\u00e7in farkl\u0131 ezme tamponlar\u0131 kullan\u0131lmaktad\u0131r. Genel olarak benim kullanmd\u0131\u011f\u0131m ezme tamponunun haz\u0131rlanmas\u0131n\u0131 anlataca\u011f\u0131m. SOD, CAT, APX, GPX, POD ve PPO enzim aktivitelerini belirlemede ayn\u0131 tamponu kullanabilirsiniz. \u00d6ncelikle 100mM (pH 7,5) k-fosfat tamponunuz haz\u0131r bulunmal\u0131d\u0131r. Nas\u0131l haz\u0131rland\u0131\u011f\u0131n\u0131 bu yaz\u0131m\u0131zda bulabilirsiniz.<\/a><\/p>\n

Kimyasallar\u0131n farkl\u0131 yap\u0131da olanlar\u0131 olabilir bu y\u00fczden molek\u00fcl a\u011f\u0131rl\u0131klar\u0131n\u0131 kontrol ederek i\u015fe ba\u015flaman\u0131z\u0131 \u00f6neririrm. 0,5gr 1 \u00f6rnek i\u00e7in 3 ml ezme tamponu kullan\u0131lmaktad\u0131r. buradan size gereken tampon kar\u0131\u015f\u0131m\u0131n\u0131 hesapl\u0131yabilirsiniz.<\/p>\n

E\u011fer farkl\u0131 mollerde kar\u0131\u015f\u0131m haz\u0131rlaman\u0131z gerekiyor ve hesaplamada s\u0131k\u0131nt\u0131 \u00e7ekiyorsan\u0131z molarite hesaplama i\u00e7in t\u0131klay\u0131n\u0131z<\/a>.<\/p>\n

Gerekenler<\/u><\/p>\n

100mM K-fosfat pH 7,5 (Haz\u0131rlan\u0131\u015f\u0131 i\u00e7in t\u0131klay\u0131n\u0131z<\/a>)<\/p>\n

0,5 mM EDTA (Molek\u00fcl A\u011f\u0131rl\u0131\u011f\u0131 : 372.24 gr\/mol)<\/p>\n

1mM askorbik asit (Molek\u00fcl A\u011f\u0131rl\u0131\u011f\u0131 : 182,08 gr\/mol) (Apx enzim \u00f6l\u00e7\u00fcm\u00fc yap\u0131lacak ise)<\/p>\n

%1-2 Polivinilpirolidon (PVP) (\u00f6zellikle POD ve PPO aktivite \u00f6l\u00e7\u00fcmlerinde eklenmeli).<\/p>\n

Haz\u0131rlan\u0131\u015f\u0131 (250 ml\u2019lik ezme tamponu i\u00e7in)<\/u><\/p>\n

1. 0,0186 gr EDTA\u2019y\u0131 250 ml 100mM K-fosfat tamponu i\u00e7erisine i\u00e7erisinde \u00e7\u00f6z\u00fcn\u00fcz.<\/p>\n

2. 0,0046 gr askorbik asit ekle.<\/p>\n

3. Son olarak %1\u2019lik istiyorsan 2,5 gr PVP tart\u0131p kar\u0131\u015f\u0131m\u0131n \u00fczerine ekleyerek manyatik kar\u0131\u015ft\u0131r\u0131c\u0131 ile iyice kar\u0131\u015ft\u0131r\u0131n\u0131z.<\/p>\n

4. 0,5 gr bitki \u00f6rne\u011finizden so\u011futulmu\u015f yada buz i\u00e7erisindeki havana koyup haz\u0131rlad\u0131\u011f\u0131n\u0131z tampondan 3 ml \u00fczerine ekleyip iyice eziniz.<\/p>\n

5. Kar\u0131\u015f\u0131m\u0131 t\u00fcplere al\u0131p 10,000x g 4 \u00b0C de 20 dk santf\u00fcr\u00fcj edin.<\/p>\n

6. S\u00fcpernatand\u0131 (\u00fcst s\u0131v\u0131) temiz bir t\u00fcpe aktar\u0131p hemen \u00f6l\u00e7\u00fcm yapacaksan\u0131z +4o<\/sup>C\u2019ye yada -20o<\/sup>C<\/p>\n

de saklay\u0131n\u0131z.<\/p>\n

Not: Katalaz \u00e7abuk bozulur. Bu y\u00fczden buzla \u00e7al\u0131\u015fmay\u0131 ihmal etmeyin. \u00c7ok fazla \u00f6rne\u011finiz varsa ayn\u0131 s\u00fcpernatant\u0131 iki yada bir ka\u00e7 t\u00fcpe da\u011f\u0131tarak devaml\u0131 olarak \u00e7\u00f6z\u00fcl\u00fcp yeniden donmas\u0131ndan dolay\u0131 olu\u015facak bozulmalar\u0131 en aza indirmi\u015f olursunuz.<\/p>\n","protected":false},"excerpt":{"rendered":"

Ba\u015flamadan \u00f6nce her zaman dedi\u011fim gibi literat\u00fcre iyice bir bak\u0131n. Baz\u0131 durumlarda farkl\u0131 bitkiler i\u00e7in farkl\u0131 ezme tamponlar\u0131 kullan\u0131lmaktad\u0131r. Genel olarak benim kullanmd\u0131\u011f\u0131m ezme tamponunun haz\u0131rlanmas\u0131n\u0131 anlataca\u011f\u0131m. SOD, CAT, APX,…<\/p>\n","protected":false},"author":1,"featured_media":1593,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"jetpack_post_was_ever_published":false,"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":true,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2},"_links_to":"","_links_to_target":""},"categories":[9],"tags":[30,26,28],"class_list":["post-248","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-genel-tr","tag-buffer","tag-enzim","tag-tampon"],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"https:\/\/orobanche.net\/wp-content\/uploads\/2018\/09\/pexels-photo-3825379.jpeg","jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/p9Bqz9-40","jetpack_likes_enabled":true,"jetpack-related-posts":[{"id":219,"url":"https:\/\/orobanche.net\/?p=219&lang=tr","url_meta":{"origin":248,"position":0},"title":"Fosfat Tamponu Nas\u0131l Haz\u0131rlan\u0131r ?","author":"graiv","date":"A\u011fustos 26, 2018","format":false,"excerpt":"\u00a0 E\u011fer literat\u00fcrde ba\u015fka bir tampon kullanman\u0131z belirtilmemi\u015f ise enzim \u00e7al\u0131\u015fmalar\u0131nda Potasyum yada Sodyum fostat tamponlar\u0131n\u0131 kullanabilirsiniz. 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Haz\u0131rlamak i\u00e7in; Dibazik potasyum fostat (K2HPO4)\u2026","rel":"","context":""Akademik Yaz\u0131lar\u0131m" i\u00e7inde","block_context":{"text":"Akademik Yaz\u0131lar\u0131m","link":"https:\/\/orobanche.net\/?cat=9&lang=tr"},"img":{"alt_text":"man in white dress shirt holding white paper","src":"https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2018\/08\/pexels-photo-6208927.jpeg?resize=350%2C200&ssl=1","width":350,"height":200,"srcset":"https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2018\/08\/pexels-photo-6208927.jpeg?resize=350%2C200&ssl=1 1x, https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2018\/08\/pexels-photo-6208927.jpeg?resize=525%2C300&ssl=1 1.5x, https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2018\/08\/pexels-photo-6208927.jpeg?resize=700%2C400&ssl=1 2x"},"classes":[]},{"id":296,"url":"https:\/\/orobanche.net\/?p=296&lang=tr","url_meta":{"origin":248,"position":1},"title":"Agaroz Jel Elektroforezi i\u00e7in tampon haz\u0131rlamak","author":"graiv","date":"Ocak 1, 2019","format":false,"excerpt":"Jel elekroforozi, negatif y\u00fckl\u00fc olan DNA par\u00e7ac\u0131klar\u0131n\u0131n agaroz jel \u00fczerinde eksi-art\u0131 kutbu boyunca hareket etmesi ve bu sayede molek\u00fcl a\u011f\u0131rl\u0131\u011f\u0131na g\u00f6re DNA par\u00e7alar\u0131n\u0131n ayr\u0131lmas\u0131nda kullan\u0131lan bir y\u00f6ntemdir (Tabi ki jel elektroforozi sadece DNA\u2019 n\u0131n ayr\u0131lmas\u0131nda kullan\u0131lmaz protein ayr\u0131lmas\u0131 gibi farkl\u0131 kullan\u0131m ama\u00e7lar\u0131 mevcuttur).\u00a0 Jel tank\u0131nda bu hareketi sa\u011flayacak bir iyonik\u2026","rel":"","context":""Akademik Yaz\u0131lar\u0131m" i\u00e7inde","block_context":{"text":"Akademik Yaz\u0131lar\u0131m","link":"https:\/\/orobanche.net\/?cat=9&lang=tr"},"img":{"alt_text":"woman in white laboratory gown","src":"https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2019\/01\/pexels-photo-3735782.jpeg?resize=350%2C200&ssl=1","width":350,"height":200,"srcset":"https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2019\/01\/pexels-photo-3735782.jpeg?resize=350%2C200&ssl=1 1x, https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2019\/01\/pexels-photo-3735782.jpeg?resize=525%2C300&ssl=1 1.5x, https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2019\/01\/pexels-photo-3735782.jpeg?resize=700%2C400&ssl=1 2x, https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2019\/01\/pexels-photo-3735782.jpeg?resize=1050%2C600&ssl=1 3x, https:\/\/i0.wp.com\/orobanche.net\/wp-content\/uploads\/2019\/01\/pexels-photo-3735782.jpeg?resize=1400%2C800&ssl=1 4x"},"classes":[]},{"id":1559,"url":"https:\/\/orobanche.net\/?p=1559&lang=tr","url_meta":{"origin":248,"position":2},"title":"Bitkilerde Prolin Analizi","author":"graiv","date":"Ocak 17, 2022","format":false,"excerpt":"Bulundurman\u0131z Gereken Malzemeler ProlinNinhidrinGlacial acetic acidEthonol (%98)Saf suEppendorf T\u00fcpBlok \u0131s\u0131t\u0131c\u0131 yada su banyosuSantrif\u00fcjmikroplateMikroplate Spektrofotometre Prolin i\u00e7eri\u011fini - \u03bcmol g-1 FW\u00a0 cinsinden hesaplayaca\u011f\u0131z. 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