| 缁嗚優鍩烘湰灞炴€?/strong> |
| 缁嗚優鍚嶇О |
HOS-LUC-PURO(浜洪鑲夌槫缁嗚優-鑽у厜绱犻叾鏍囪) |
| 缁嗚優鍒О |
HOS-LUC锛涗汉楠ㄨ倝鐦ょ粏鑳炴牚-鑽у厜绱犻叾鏍囪锛涗汉楠ㄨ倝鐦ょ粏鑳炴牚-LUC-PURO |
| 绉嶅睘鏉ユ簮 |
浜裹/span> |
| 缁勭粐鏉ユ簮 |
楠?/span> |
| 鐢熼暱鐗规€?/span> |
璐村鐢熼暱 |
| 缁嗚優褰㈡€?/span> |
涓婄毊鏍风粏鑳炲拰鎴愮氦缁存牱缁嗚優鍧囨湁 |
| 缁嗚優浠ｆ暟 |
p3-p8 |
| 鑳屾櫙浠嬬粛 |
Luciferase HOS缁嗚優绋冲畾琛ㄨ揪钀ょ伀铏崸鍏夌礌閰躲€傝缁嗚優鏍€х姸绋冲畾锛屽煿鍏绘椂涓嶉渶瑕佹坊鍔犳姉鐢熺礌缁存寔銆傚彲鐢ㄤ綔钀ょ伀铏崸鍏夌礌閰舵椿鎬ф娴嬩腑鐨勯槼鎬у鐓э紝涔熷彲鐢ㄤ簬娲讳綋鍔ㄧ墿鎴愬儚瀹為獙銆侟/span> 璇ョ粏鑳為€氳繃鎱㈢梾姣掕浆鏌撶殑鏂瑰紡鎼哄甫Luc鍩哄洜銆侟/span> HOS缁嗚優鏄粠涓€鍚?3宀佺殑鐧戒汉濂虫€х殑楠ㄨ倝鐦ょ粍缁囦腑鍒嗙寤虹珛鐨勶紱HOS缁嗚優琛ㄧ幇涓烘墎骞冲舰鎬併€佷綆楗卞拰瀵嗗害銆佽蒋鐞艰剛涓殑浣庡厠闅嗗舰鎴愮巼鍜屽鍖栧悎鐗╁強鐥呮瘨鎰熸煋鐨勬晱鎰熸€с€侟/span> |
| puro鑽瓫娴撳害 |
HOS-LUC缁嗚優puro鑽瓫娴撳害涓?. 0ug/ml锛屽煿鍏昏繃绋嬩腑鍙笉鐢ㄥ啀娣诲姞puro锛屽鑻ユ媴蹇冩姉鎬ч殢鐫€浼犱唬鏃堕棿闄嶄綆锛屽彲瀹氭湡鐢?.5ug/ml娴撳害puro缁存寔 |
| 鐢熺墿瀹夊叏绛夌骇 |
1 |
| Luciferase娲绘€ф娴婞/span> |
|
| STR閴村畾鍥剧墖 |
|
| 缁嗚優瑙勬牸 |
1x106cells/T25鍩瑰吇鐡舵垨鑰?mL鍐诲瓨绠″寘瑁匋/span> |
| 鏀師浣撴娴婞/span> |
鏃燑/span> |
| 淇濊棌鏈烘瀯 |
ATCC; CRL-1543 ECACC; 87070202 |
| 鍩瑰吇鍩裹/span> |
90%MEM+10% FBS+PS |
| 鍩瑰吇鏉′欢 |
姘旂浉锛?5%绌烘皵+5%浜屾哀鍖栫⒊锛涙俯搴︼細37鈩傸/span> |
| 鍐诲瓨鏉′欢 |
鏃犺娓呭喕瀛樻恫锛屾恫姘偍瀛楛/span> |
| 缁嗚優璐ф湡 |
鐜拌揣锛?鍛ㄥ乏鍙颤/span> |
| 杩愯緭鏂瑰紡 |
澶嶈嫃鍙戣揣(T25鐡跺厤杩愯緭璐圭敤)/ 鍐诲瓨鍙戣揣(闇€鍔犲共鍐拌繍杈撹垂鐢? 椤轰赴蹇€扅/span> |
| 渚涘簲闄愬埗 |
浠呴檺浜庣瀛︾爺绌讹紝缁濅笉鍙綔涓哄姩鐗╂垨浜虹被鐤剧梾鐨勫尰鐤椾骇鍝佷娇鐢?/span> |
| 鐗瑰埆璇存槑 |
浠ヤ笅缁嗚優鍩瑰吇鍐诲瓨澶勭悊浠呬緵鍙傝€冿紝鍏蜂綋鎿嶄綔姝ラ浠ラ殢璐т骇鍝佽鏄庝功涓轰富 |
| 缁嗚優鍩瑰吇鎿嶄綔 |
| 鏀惰揣鏂瑰紡 |
T25鐡迭/strong> |
鍐诲瓨绠狘/strong> |
| 鏀惰揣澶勭悊 |
瑙傚療濂界粏鑳炵姸鎬佸悗锛?5%閰掔簿娑堟瘨鐡跺锛屽皢T25鐡剁疆浜?7搴﹀煿鍏荤鏀剧疆2-4h锛屼互渚跨ǔ瀹氱粏鑳炵姸鎬?/span> |
鏀跺埌缁嗚優鍚庯紝闇€绔嬪嵆杞叆娑叉爱鍐诲瓨鎴栫洿鎺ュ鑻廃/span> |
| 浼犱唬瀵嗗害 |
缁嗚優瀵嗗害杈?0%-90%锛屽嵆鍙繘琛屼紶浠ｅ煿鍏狐/span> |
绗?澶╂崲娑插苟妫€鏌ョ粏鑳炲瘑搴?/span> |
| 浼犱唬姣斾緥 |
鍒濇浼犱唬寤鸿1锛?浼犱唬
1:2浼犱唬灏辨槸1涓猅25鐡朵紶2涓猅25鐡舵垨鑰?涓?cm鐨裤€備笉鏄?涓猅25鐡朵紶2涓?0cm鐨军/strong> |
涓€绠＄粏鑳炲缓璁帴绉嶅埌10cm鍩瑰吇鐨挎垨鑰匱25鐡迭/span> |
| 浼犱唬鏂规硶 |
a銆佸純鍘诲煿鍏讳笂娓咃紝鐢ㄤ笉鍚挋銆侀晛绂诲瓙鐨凱BS娑︽礂缁嗚優1-2娆°€
b銆佸姞1 mL娑堝寲娑?0.25%Trypsin-0.53mM EDTA)浜庡煿鍏荤摱涓紝浣挎秷鍖栨恫娴告鼎鎵€鏈夌粏鑳烇紝灏嗗煿鍏荤摱缃簬37鈩冨煿鍏荤涓秷鍖?-2 min锛岀劧鍚庡湪鏄惧井闀滀笅瑙傚療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2-3ml瀹屽叏鍩瑰吇鍩虹粓姝㈡秷鍖栥€傝交杞绘墦鍖€鍚庤鍏ユ棤鑿岀蹇冪涓紝1000 rpm绂诲績5 min锛屽純鍘讳笂娓呮恫锛岃ˉ鍔?-2 mL鍩瑰吇娑插悗鍚瑰寑銆
c銆佸皢缁嗚優鎮恫鎸?:2姣斾緥鍒嗗埌鏂扮殑鍚? mL鍩瑰吇鍩虹殑鏂扮毧涓垨鑰呯摱涓紝缃簬鍩瑰吇绠变腑鍩瑰吇銆侟/span> |
灏嗗惈鏈? mL缁嗚優鎮恫鐨勫喕瀛樼鍦?7鈩冩按娴翠腑杩呴€熸憞鏅冭В鍐伙紝鍔? mL鍩瑰吇鍩烘贩鍚堝潎鍖€銆傚湪1000 rpm鏉′欢涓嬬蹇? min锛屽純鍘讳笂娓呮恫锛屽姞1-2 mL鍩瑰吇鍩哄悗鍚瑰寑銆傜劧鍚庡皢鎵€鏈夌粏鑳炴偓娑插姞鍏ュ惈閫傞噺鍩瑰吇鍩虹殑鍩瑰吇鐡朵腑鍩瑰吇杩囧(鎴栧皢缁嗚優鎮恫鍔犲叆10 cm鐨夸腑锛屽姞鍏ョ害8 mL鍩瑰吇鍩猴紝鍩瑰吇杩囧)銆傜2澶╂崲娑插苟妫€鏌ョ粏鑳炲瘑搴︺€侟/span> |
| 娉ㄦ剰浜嬮」 |
1.杩愯緭鐢ㄧ殑鍩瑰吇鍩?鐏屾恫鍩瑰吇鍩?涓嶈兘鍐嶇敤鏉ュ煿鍏荤粏鑳烇紝璇锋崲鐢ㄦ寜鐓ц鏄庝功缁嗚優鍩瑰吇鏉′欢鏂伴厤鍒剁殑瀹屽叏鍩瑰吇鍩烘潵鍩瑰吇缁嗚優銆侟/strong> 2.鍥犺繍杈撻棶棰橈紝閮ㄥ垎缁嗚優鐢变簬娓╁害鍙樺寲鍙婂墽鐑堢鎾炴浜＄牬纰庡舰鎴愮鐗囷紝鏄甯哥幇璞°€侟/span> |
1.鏀惰揣鏃惰嫢鍙戠幇骞插啺鍖栧畬锛屾鏌ュ喕瀛樼鏄惁铻嶅寲锛岃嫢宸茶瀺鍖栭渶鐩存帴绂诲績缁嗚優鎺ョ瑙傚療锛岃嫢鏈瀺鍖栧彲浠ュ皢缁嗚優鎸夋甯告楠や繚瀛橈紱
2.涓轰繚璇佺粏鑳炵殑楂樺瓨娲荤巼锛屾敹鍒颁骇鍝佸悗锛岃绔嬪嵆瑙ｅ喕澶嶈嫃缁嗚優銆侟/strong> |
| 鍒拌揣椤荤煡 |
1.鏀跺埌缁嗚優鍚庯紝棣栧厛瑙傚療骞舵媿鐓ц褰曠粏鑳炵摱鏄惁瀹屽ソ锛屽煿鍏绘恫鏄惁鏈夋紡娑层€佹祽娴婄瓑鐜拌薄锛屽共鍐拌繍杈撶殑缁嗚優妫€鏌ュ共鍐版槸鍚﹀畬鍏ㄦ尌鍙戯紝缁嗚優鏄惁瑙ｅ喕锛岃嫢鏈変笂杩扮幇璞″彂鐢熻鍙婃椂鍜屾垜浠仈绯汇€
2.闈欑疆瀹屾垚鍚庯紝鍙栧嚭缁嗚優鍩瑰吇鐡讹紝闀滄銆佹媿鐓?褰撳ぉ浠ュ強绗 2,3 澶╄鎷嶇収)锛岃褰曠粏鑳炵姸鎬 (鎵€鎷嶇収鐗囧皢浣滀负鍚庣画鏈嶅姟渚濇嵁)锛涘缓璁粏鑳炰紶浠ｅ煿鍏诲悗锛屽畾鏈熸媿鐓с€佽褰曠粏鑳炵敓闀跨姸鎬併€侟/span> 3.鐢变簬杩愯緭鐨勫師鍥狅紝涓埆鏁忔劅缁嗚優浼氬嚭鐜颁笉绋冲畾鐨勬儏鍐碉紝璇峰強鏃跺拰鎴戜滑鑱旂郴锛屽憡鐭ョ粏鑳炵殑鍏蜂綋鎯呭喌锛屼互渚挎垜浠殑鎶€鏈汉鍛樿窡韪洖璁跨洿鑷抽棶棰樿В鍐炽€侟br/>4.浠旂粏闃呰缁嗚優璇存槑涔︼紝浜嗚В缁嗚優鐩稿叧淇℃伅锛屽缁嗚優褰㈡€併€佹墍鐢ㄥ煿鍏诲熀銆佽娓呮瘮渚嬨€佹墍闇€缁嗚優鍥犲瓙绛夛紝纭繚缁嗚優鍩瑰吇鏉′欢涓€鑷达紝鑻ョ敱浜庡煿鍏绘潯浠朵笉涓€鑷磋€屽鑷寸粏鑳炲嚭鐜伴棶棰橈紝璐ｄ换鐢卞鎴疯嚜琛屾壙鎷呫€侟/strong> |
| 澶囨敞锛氬鎴峰湪缁嗚優鍩瑰吇杩囩▼涓紝鏈変换浣曟妧鏈棶棰樺彲浠ユ嫧鎵撴妧鏈湇鍔＄數璇濄€侟/span> |
| 缁嗚優鍐诲瓨鎿嶄綔 |
| 鍐诲瓨娑查厤鏂更/span> |
鏃犺娓呭喕瀛樻恫锛屾恫姘偍瀛楛/span> |
| 缁嗚優瀵嗗害 |
寰呯粏鑳炵敓闀跨姸鎬佽壇濂芥椂锛屽彲杩涜缁嗚優鍐诲瓨銆備笅闈 T25 鐡朵负渚婞/span> |
| 鍐诲瓨鏂规硶 |
a銆佹敹闆嗙粏鑳炲強缁嗚優鍩瑰吇娑诧紝瑁呭叆鏃犺弻绂诲績绠′腑锛?000 rpm鏉′欢涓嬬蹇? min锛屽純鍘讳笂娓呮恫锛岀敤PBS娓呮礂涓€閬嶏紝寮冨敖PBS锛屽姞1 mL琛€娓呴噸鎮粏鑳烇紝杩涜缁嗚優璁℃暟銆侟br/>b銆佹牴鎹粏鑳炴暟閲忓姞鍏ユ棤琛€娓呯粏鑳炲喕瀛樻恫锛屼娇缁嗚優瀵嗗害5x106锝?x107/mL锛岃交杞绘贩鍖€锛屾瘡鏀喕瀛樼鍐诲瓨1mL缁嗚優鎮恫锛屾敞鎰忓喕瀛樼鍋氬ソ鏍囪瘑銆
c銆佸皢鍐诲瓨绠℃斁鍏?80鈩冨啺绠憋紝24 h鍚庤浆鍏ユ恫姘亴鍌ㄥ瓨銆傝褰曞喕瀛樼浣嶇疆浠ヤ究涓嬫鎷垮彇銆侟/span> |
| 娉ㄦ剰浜嬮」 |
鍐诲瓨缁嗚優杞叆娑叉爱鍚庡強鏃跺鑻忎竴绠℃鏌ョ粏鑳炲喕瀛樻椿鎬э紝鑻ユ湁寮傚父锛屽強鏃惰皟鏁村疄楠屾柟妗圏/span> |
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