缁嗚優鐨勭绫伙細

绗楃锛欬/span>

HCT116-LUC鑽у厜鏍囪缁嗚優鏍?/span>鏄喕瀛樹骇鍝侊紝鐢辨恫姘洿鎺ユ嬁鍑猴紝骞插啺杩愯緭缁欏鎴枫€備竴鑸笉鎺ㄨ崘杩欑锛屼篃杈冨皯浣跨敤杩欑鏂瑰紡锛屽洜涓哄鑻忔墜娉曞浜庣粏鑳炵姸鎬佸奖鍝嶈緝澶э紝涓€鏃︾粏鑳炵姸鎬佷笉濂斤紝寰堥毦璇存槸缁嗚優鑷韩涓嶈锛岃繕鏄鑻忔病鎿嶄綔濂斤紱鍙﹀娓╁害瀵圭粏鑳炪€佸熀鍥犲姛鑳界姸鎬佸奖鍝嶅緢澶э紝涔熶笉鏄粏鑳炲偍瀛樼殑zui濂芥柟寮忥紝蹇€掓椂闂翠篃寰堥毦鎺у埗锛屽绉嶅洜绱犲奖鍝嶄骇鍝佺殑璐ㄩ噺锛涘共鍐拌繍杈撻渶瑕侀澶栨坊鍔?00鍏冪殑杩愯垂(椤轰赴)銆侟/span>

绗簩绉嶏細

鏄垜浠鑻忓ソ缁嗚優锛孅/span>QC閫氳繃鍚庯紝T-25鐏屾弧鍩瑰吇鍩哄父娓╄繍杈撶粰瀹㈡埛锛屾帓闄ゅ鑻忛€犳垚褰卞搷锛屽父娓╄繍杈撲篃瀵圭粏鑳炲奖鍝嶄篃涓嶅ぇ锛屽彧闇€鍔?5鍏冭繍璐?椤轰赴)銆侟/span>

璐ㄩ噺淇濊瘉锛氭垜浠殑缁嗚優鏍潵婧愪簬ATCC銆丒CACC銆丏SMZ銆丣CRB绛夛紝璐拱鍒拌揣鍚庯紝鐢辨垜浠疄楠屽鎵╁銆佸喕瀛橈紝鍐诲瓨鐨勪骇鍝佸叏閮ㄧ粡杩嘠C妫€娴嬶紝100%杩涘彛鏉ユ簮锛?00%淇濊瘉5浠ｄ互鍐咃紝娲诲姏>95%锛屾棤缁嗚弻銆佺湡鑿屻€佹敮鍘熶綋姹℃煋銆傜殑鍐诲瓨鏄粏鑳炲煿鍏荤殑鍩烘湰鎿嶄綔锛屼笉鍚岀殑瀹為獙瀹ら噰鐢ㄧ殑鏂规硶鐣ユ湁宸紓锛屼絾鏄兘浼氶伒寰參鍐婚€熸憾鐨勫畻鏃ㄣ€侟/span>
鐜拌揣渚涘簲锛屼环鏍间紭鎯犮€佸疄楠屾晥鏋滃ソ锛屾垜鍙镐负鎮ㄦ彁渚涘厤璐逛唬娴嬫湇鍔★紝鍚屾椂鎻愪緵鏈€鏂颁环鏍笺€佽鏄庝功銆佽鏍笺€佺敤閫斻€佸疄楠屽師鐞嗙瓑鐩稿叧鎿嶄綔璇存槑銆侟/span>

浜у搧鍚嶇О	HCT116-LUC鑽у厜鏍囪缁嗚優鏍?/span>
浜у搧绠€绉?/span>	HCT116-LUC
璐у彿	GOY-N0151

銆愬煿鍏绘寚鍗椼€慄/span>

瀵逛簬璐村缁嗚優锛屼紶浠ｅ彲鍙傝€冧互涓嬫柟娉曪細

1. 寮冨幓鍩瑰吇涓婃竻锛岀敤涓嶅惈閽欍€侀晛绂诲瓙鐨凱BS娑︽礂缁嗚優1-2娆°€侟/span>

2. 鍔?ml娑堝寲娑?0.25%Trypsin-0.53mM EDTA)浜庡煿鍏荤摱涓紝缃簬37鈩冨煿鍏荤涓秷鍖?-2鍒嗛挓锛岀劧鍚庡湪鏄惧井闀滀笅瑙傚療鈪″瀷鑲烘场涓婄毊缁嗚優|缁嗚優娑堝寲鎯呭喌锛岃嫢缁嗚優澶ч儴鍒嗗彉鍦嗗苟鑴辫惤锛岃繀閫熸嬁鍥炴搷浣滃彴锛岃交鏁插嚑涓嬪煿鍏荤摱鍚庡姞灏戦噺鍩瑰吇鍩虹粓姝㈡秷鍖栥€侟/span>

3. 鎸?-8ml/鐡惰ˉ鍔犲煿鍏诲熀锛岃交杞绘墦鍖€鍚庡惛鍑猴紝鍦?000RPM鏉′欢涓嬬蹇?鍒嗛挓锛屽純鍘讳笂娓呮恫锛岃ˉ鍔?-2mL鍩瑰吇娑插悗鍚瑰寑銆侟/span>

4. 灏嗙粏鑳炴偓娑叉寜1锛?鍒?锛?鐨勬瘮渚嬪垎鍒版柊鐨勫惈8ml鍩瑰吇鍩虹殑鏂扮毧涓垨鑰呯摱涓€侟/span>

缁嗚優鍩瑰吇鐨勪紭鐐癸細

1.鐮旂┒鐨勫璞℃槸娲荤粏鑳濣/span>

鍦ㄥ疄楠岃繃绋嬩腑锛屾牴鎹姹傚彲濮嬬粓淇濇寔缁嗚優娲诲姏锛屽苟鍙暱鏃堕棿鐩戞帶銆佹娴嬬敋鑷冲畾閲忚瘎浼颁竴閮ㄥ垎娲荤粏鑳炵殑鎯呭喌锛屽寘鎷椿缁嗚優鐨勫舰鎬併€佺粨鏋勩€佺敓鍛芥椿鍔ㄧ瓑銆侟/span>

2.鐮旂┒鏉′欢鍙互浜轰负鎺у埗

pH銆佹俯搴︺€佹哀姘斻€佷簩姘у寲纰炽€佸紶鍔涚瓑鐗╃悊鍖栧鐨勬潯浠讹紝鍙互鏍规嵁瀹為檯闇€瑕佽繘琛屼汉涓虹殑鎺у埗锛屽悓鏃讹紝鍙互鏂藉姞鍖栧銆佺墿鐞嗐€佺敓鐗╃瓑鍥犵礌浣滀负鏉′欢鑰岃繘琛屽疄楠岃瀵燂紝杩欎簺鍥犵礌鍚屾牱鍙互澶勪簬涓ユ牸鎺у埗涔嬩笅銆侟/span>

3.鐮旂┒鐨勬牱鏈彲浠ヨ揪鍒版瘮杈冨潎涓€鎬?/span>

閫氳繃缁嗚優鍩瑰吇涓€瀹氫唬鏁板悗锛屾墍寰楀埌鐨勭粏鑳炵郴鍒欏彲浠ヨ揪鍒板潎涓€鎬ц€屽睘浜庡悓涓€绫诲瀷鐨勭粏鑳烇紝闇€瑕佹椂锛屽彲閲囩敤鍏嬮殕鍖栫瓑鏂规硶浣跨粏鑳炶揪鍒扮函鍖栥€侟/span>

4.鐮旂┒鍐呭渚夸簬瑙傚療銆佹娴嬪拰璁板綍

閲囩敤鍚勭鐮旂┒鎶€鏈細濡傚€掔疆鐢熺墿鏄惧井闀溿€佽崸鍏夋樉寰暅銆佺數瀛愭樉寰暅銆佹祦寮忕粏鑳炴湳銆佹縺鍏夊叡鐒︽樉寰暅銆佸厤鐤粍缁囧寲瀛︺€佸師浣嶆潅浜ゃ€佸悓浣嶇礌鏍囪绛夊悇绉嶄华鍣ㄨ澶囥€侟/span>
灏忛紶鎴愮氦缁寸粏鑳炲畬鍏ㄥ煿鍏诲熀Zika virus鐗逛环渚涘簲96T

灏忛紶琛ㄧ毊瑙掑寲缁嗚優瀹屽叏鍩瑰吇鍩裹/span>Cytochrome 5A鐜拌揣渚涘簲96T

灏忛紶鐪熺毊鎴愮氦缁寸粏鑳炲畬鍏ㄥ煿鍏诲熀Glutamate--cysteine ligase catalytic subunit鐗逛环淇冮攢96T

灏忛紶杞缁嗚優瀹屽叏鍩瑰吇鍩裹/span>Glutamate--cysteine ligase modifier subunit涓婃捣鐩撮攢96T

灏忛紶鐮撮缁嗚優瀹屽叏鍩瑰吇鍩裹/span>glutathione synthetase鐗逛环渚涘簲96T

灏忛紶鐨偆鑲ュぇ缁嗚優瀹屽叏鍩瑰吇鍩裹/span>spermidine synthase鐜拌揣渚涘簲96T

灏忛紶鍓嶈剛鑲粏鑳炲畬鍏ㄥ煿鍏诲熀TGF-beta-activated kinase 1 and MAP3K7-binding protein 3鐗逛环淇冮攢96T

灏忛紶鎴愰缁嗚優瀹屽叏鍩瑰吇鍩裹/span>Phytohaemagglutinin涓婃捣鐩撮攢96T

灏忛紶鍏宠妭杞缁嗚優瀹屽叏鍩瑰吇鍩裹/span>Varicella-zoster virus鐗逛环渚涘簲96T

灏忛紶鑳庡効鐪熺毊鎴愮氦缁寸粏鑳炲畬鍏ㄥ煿鍏诲熀Lebtospira Ab鐜拌揣渚涘簲96T
HCT116-LUC鑽у厜鏍囪缁嗚優鏍?/span>4--3-姘熻媱鍩烘捍

4--3-姘熻媱鍩烘捍

1-姝?涓佸熀-4-鑻?/span>

4-涓佸熀鑻敳

4-涓佸熀鑻敳

4-涓佸熀鑻敳

鑻哀涔欑敳閰?/span>

5-涔欓叞鍩哄櫥鍚?2-纭稽/span>

5-涔欓叞鍩哄櫥鍚?2-纭稽/span>

5-涔欓叞鍩哄櫥鍚?2-纭稽/span>
銆愮粏鑳炲喕瀛樸€慄/span>

浜у搧鍚嶇О寰呯粏鑳炵敓闀跨姸鎬佽壇濂芥椂锛屽彲杩涜293/HEK-293缁嗚優|缁嗚優鍐诲瓨銆傝创澹佺粏鑳炲喕瀛樻椂锛屽純鍘诲煿鍏诲熀鍚庡姞鍏ュ皯閲忚儼閰讹紝缁嗚優鍙樺渾鑴辫惤鍚庯紝鍔犲叆绾?ml鍚娓呯殑鍩瑰吇鍩哄悗鍔犲叆鍐诲瓨绠′腑锛屽啀娣诲姞10%DMSO鍚庤繘琛屽喕瀛樸€傛偓娴粏鑳炲喕瀛樻椂锛屽簲灏嗙粏鑳炴敹闆嗭紝1000RPM鏉′欢涓嬬蹇?鍒嗛挓锛屽皯閲忎繚瀛樹笂娓呮恫(闃叉缁嗚優鍚歌蛋)锛屽姞鍏ラ儴鍒嗘柊椴滃煿鍏诲熀锛屽姞鍏ュ埌鍐诲瓨绠′腑锛屽湪鍐诲瓨绠′腑鍔犲叆10%DMSO鍚庤繘琛屽喕瀛樸€侟/span>

銆愬鑻忕殑鍘熷垯銆慄/span>

蹇€熻瀺鍖栵細蹇呴』灏嗗喕瀛樺湪-196鈩冩恫姘腑鐨勭粏鑳炲揩閫熻瀺鍖栬嚦37鈩冿紝浣跨粏鑳炲鍐诲瓨鏃剁殑鍐版櫠杩呴€熻瀺鍖栵紝閬垮厤鍐版櫠缂撴參铻嶅寲鏃惰繘鍏ョ粏鑳炲舰鎴愬啀缁撴櫠锛屽缁嗚優閫犳垚鎹熷銆傚鏃︾粏鑳炲簱鍏ㄥ浗鍚勫湴鍧囧彲鍙戣揣锛屽叏鍥界粺涓€浠锋牸锛屾湰鍏徃鍑哄敭鐨勬墍鏈夌粏鑳炰粎渚涚鐮斾娇鐢ㄣ€侟/span>