鐗规€э細

COC1/DDP浜哄嵉宸㈢檶椤洪搨鑰愯嵂鏍?/span>瀹夊叏鎬э細鎵€鏈夎偪鐦ゅ拰鐥呮瘨杞煋鐨勭粏鑳炲潎瑙嗕负鏈夋綔鍦ㄧ殑鐢熺墿鍗卞鎬э紝蹇呴』鍦ㄤ簩绾х敓鐗╁畨鍏ㄥ彴鍐呮搷浣滐紝骞惰娉ㄦ剰闃叉姢锛屾墍鏈夊簾娑插強鎺ヨЕ杩囨缁嗚優鐨勫櫒鐨块渶楂樺帇鐏弻鍚庢柟鑳戒涪寮冦€傛敹鍒扮粏鑳炲悗锛屽湪鍊掔疆闀滀笅濂芥槸鍦?/span>4X鐗╅暅)瑙傚療鏁翠釜缁嗚優鐢熼暱鎯呭喌

锛堜竴锛夊鏋滅粏鑳炴湭闀挎弧锛岀敤75%閰掔簿鍠锋磼鏁翠釜鐡舵秷姣掑悗鏀惧埌瓒呰弻鍙板唴锛屼弗鏍兼棤鑿屾搷浣滐紝鎵撳紑缁嗚優鍩瑰吇鐡讹紝鍚稿嚭鍩瑰吇娑诧紝鎹 10ml鏂伴矞鍩瑰吇娑插悗缁х画鍩瑰吇銆侟/span>

(浜?濡傛灉缁嗚優宸查暱婊★紝鍗冲彲杩涜浼犱唬鍩瑰吇銆傚叿浣撴楠ゅ涓嬶細

1. 寮冨幓鍩瑰吇娑诧紝鐢≒BS锛堜笉鍚挋,闀佺瀛愶級娲?-2娆°€侟/span>

2. 鍔?ml娑堝寲娑诧紙0.25%Trypsin-0.53mM EDTA锛変簬鍩瑰吇鐡朵腑锛屽€掓斁浜?7鈩冨煿鍏荤1-3鍒嗛挓棰勭儹锛屼箣鍚庣炕杞煿鍏荤摱10-30绉掑悗锛屽湪鍊掔疆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儴鍒嗗彉鍦嗭紝杩呴€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瀹屽叏鍩瑰吇鍩虹粓姝㈡秷鍖栥€侟/span>

3. 鎸?-8ml/鐡惰ˉ鍔犲畬鍏ㄥ煿鍏诲熀锛岃交杞绘墦鍖€鍚庡惛鍑轰竴鍗婏紝鍒嗗埌鏂扮殑鍩瑰吇鐡朵腑銆侟/span>

浜у搧鍚嶇О	浜у搧绠€绉?/span>	璐у彿
COC1/DDP浜哄嵉宸㈢檶椤洪搨鑰愯嵂鏍?/span>	COC1/DDP	GOY-N0003

鐗规€э細

瀹夊叏鎬э細鎵€鏈夎偪鐦ゅ拰鐥呮瘨杞煋鐨勭粏鑳炲潎瑙嗕负鏈夋綔鍦ㄧ殑鐢熺墿鍗卞鎬э紝蹇呴』鍦ㄤ簩绾х敓鐗╁畨鍏ㄥ彴鍐呮搷浣滐紝骞惰娉ㄦ剰闃叉姢锛屾墍鏈夊簾娑插強鎺ヨЕ杩囨缁嗚優鐨勫櫒鐨块渶楂樺帇鐏弻鍚庢柟鑳戒涪寮冦€傛敹鍒扮粏鑳炲悗锛屽湪鍊掔疆闀滀笅濂芥槸鍦?/span>4X鐗╅暅)瑙傚療鏁翠釜缁嗚優鐢熼暱鎯呭喌

锛堜竴锛夊鏋滅粏鑳炴湭闀挎弧锛岀敤75%閰掔簿鍠锋磼鏁翠釜鐡舵秷姣掑悗鏀惧埌瓒呰弻鍙板唴锛屼弗鏍兼棤鑿屾搷浣滐紝鎵撳紑缁嗚優鍩瑰吇鐡讹紝鍚稿嚭鍩瑰吇娑诧紝鎹 10ml鏂伴矞鍩瑰吇娑插悗缁х画鍩瑰吇銆侟/span>

(浜?濡傛灉缁嗚優宸查暱婊★紝鍗冲彲杩涜浼犱唬鍩瑰吇銆傚叿浣撴楠ゅ涓嬶細

1. 寮冨幓鍩瑰吇娑诧紝鐢≒BS锛堜笉鍚挋,闀佺瀛愶級娲?-2娆°€侟/span>

2. 鍔?ml娑堝寲娑诧紙0.25%Trypsin-0.53mM EDTA锛変簬鍩瑰吇鐡朵腑锛屽€掓斁浜?7鈩冨煿鍏荤1-3鍒嗛挓棰勭儹锛屼箣鍚庣炕杞煿鍏荤摱10-30绉掑悗锛屽湪鍊掔疆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儴鍒嗗彉鍦嗭紝杩呴€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瀹屽叏鍩瑰吇鍩虹粓姝㈡秷鍖栥€侟/span>

3. 鎸?-8ml/鐡惰ˉ鍔犲畬鍏ㄥ煿鍏诲熀锛岃交杞绘墦鍖€鍚庡惛鍑轰竴鍗婏紝鍒嗗埌鏂扮殑鍩瑰吇鐡朵腑銆侟/span>

缁嗚優鐗规€э細

1)缁勭粐鏉ユ簮浜庡疄楠屽姩鐗╃殑姝ｅ父鐪肩粍缁囥€侟/span>

2)缁嗚優閴村畾锛氱粏鑳炶铔嬬櫧-18(CK-18)鍏嶇柅鑽у厜鏌撹壊涓洪槼鎬с€侟/span>

3)缁忛壌瀹氱粏鑳炵函搴﹂珮浜?0%銆侟/span>

4)涓嶅惈鏈 HIV-1銆 HBV銆丠CV銆佹敮鍘熶綋銆佺粏鑿屻€侀叺姣嶅拰鐪熻弻銆侟/span>

5)缁嗚優鐢熼暱鏂瑰紡锛氫笂鐨牱锛屼笉瑙勫垯缁嗚優锛岃创澹佸煿鍏汇€侟/span>

娉ㄦ剰浜嬮」锛欬/span>

1銆佽瀵熺粏鑳瀦ui濂藉湪浣庡€嶉暅锛?鎴?X鐗╅暅)涓嬭繘琛岋紝鍚﹀垯涓嶈兘鍑嗙‘鍒ゆ柇缁嗚優鐨勪紶浠ｅ瘑搴︺€傜湅缁嗚優鐨勫舰鎬佽鍦?0X鎴?0X鐗╅暅涓嬨€侟/span>

2銆佺摱涓繍杈撳煿鍏诲熀涓嶈兘閲嶅鍐嶇敤,璇锋崲鐢ㄥ姞鍙屾姉鐨勬柊鍩瑰吇鍩猴紝缁嗚優鍐诲瓨鍚?鍩瑰吇鍩轰腑鍙笉鍔犱换浣曟姉鐢熺礌銆侟/span>

3銆佹湁浜涚粏鑳炶创澹佷笉鐗紝濡傚彂鐜拌创澹佺粏鑳炴湁鑴辫惤锛屽彲绂诲績鍚规墦鍚庢帴绉嶅埌鏂扮摱鍐呫€侟/span>

4銆佹敹鍒扮粏鑳炲悗锛岃嫢鍙戠幇鍩瑰吇鐡剁牬鎹熴€佹湁娑叉孩鍑哄強缁嗚優鏈夋薄鏌擄紝璇峰強鏃朵笌鎴戜滑銆侟/span>

3,4,5-涓夋盁鑻箼

3,4,5-涓夋盁鑻箼

3-姘ㄥ熀-5-姘熷悺鍟迭/span>

3-姘ㄥ熀-5-姘熷悺鍟迭/span>

(S)-DTBM-SEGPHOS

(S)-DTBM-SEGPHOS

2,3,4-涓夌敳姘у熀鑻敳

2,3,4-涓夌敳姘у熀鑻敳

2,3,4-涓夌敳姘у熀鑻敳

姘у寲閽滁/span>

Prostaglandin F2伪,PGF2伪 ELISA KitHPLC鈮?8% 4瀛擬IP-5 ELISA Kit

Toll-like receptor 4,TLR4 ELISA KitHPLC鈮?8%6MIP-3尾 ELISA kit

soluble myosin heavy chain 1,sMHC-1 ELISA KitHPLC鈮?7%100锝?000MIP-3伪 ELISA kit

soluble myosin heavy chain 2,sMHC-2 ELISA KitHPLC鈮?8%4487macrophage inflammatory protein 2,MIP-2 ELISA Kit

Mucin-5 subtype AC,MUC5AC ELISA KitHPLC鈮?8%96 wells,0.2MIP-1尾 ELISA kit

epidermal growth factor receptor,EGFR ELISA KitHPLC鈮?8%600.0mlMIP-1伪 ELISA kit

transthyretin,TTR ELISA KitHPLC鈮?8%MDC ELISA kit

瀹為獙鎶ュ憡锛欬/span>

涓€銆佸垎绂讳笌鍩瑰吇锛欬/span>

1銆佹棤鑿屾潯浠朵笅锛屽彇鍑?-3d 榫凷D澶ч紶蹇冩埧缁勭粐锛岀劧鍚庣敤PBS灏嗘缁勭粐鍧楁竻娲?娆★紝鏈€鍚庡皢缁勭粐鍓垚1mm3宸﹀彸澶у皬锛汓/span>

2銆佸線缁勭粐鍧椾腑鍔犲叆4 mL閰舵秷鍖栨恫锛?.1% 鑳伴叾鍜?.1% I鍨嬭兌鍘熼叾锛夛紝娣锋偓10s锛岀疆37鈩冩潯浠朵笅娑堝寲10min锛屼箣鍚庣敤婊寸鍚规墦鍒舵垚鍗曠粏鑳炴偓娑诧紝鑷劧娌夋穩骞舵敹闆嗕笂娓咃紝鐢ㄥ惈10% FBS鍩瑰吇鍩虹粓姝㈡秷鍖栧悗4鈩冩斁缃紱

3銆佸墿涓嬬殑缁勭粐鍐嶅姞鍏?锝?mL閰舵秷鍖栨恫锛屾贩鎮?0s锛岀疆37鈩冩秷鍖?0 min鍚庯紝鎸変笂杩版柟娉曟敹闆嗕笂娓呭苟缁堟娑堝寲鍚?鈩冩斁缃紝閲嶅姝ゆ楠?-3娆★紝鐩磋嚦缁勭粐瀹屽叏琚秷鍖栵紱

4銆佺敤200鐩笉閿堥挗绛涚綉杩囨护缁嗚優娑堝寲娑诧紝1200r/min 绂诲績10min锛屽純鍘讳笂娓咃紝娌夋穩缁嗚優鐢ㄥ惈鏈?0锛 FBS DMEM/F12鍩瑰吇鍩烘贩鎮紝鎺ョ浜?5cm2鍩瑰吇鐡讹紝鏀剧疆浜?7鈩 锛?锛匔O2 鍩瑰吇绠变腑鍩瑰吇锛汓/span>

5銆佸樊閫熻创澹?h鍚庯紝鍚稿嚭鍩瑰吇鍩猴紝鎸夊疄楠岄渶瑕佹帴绉嶄簬6瀛旀澘涓户缁煿鍏伙紱

浜屻€佸厤鐤崸鍏夐壌瀹氾細

1銆佸緟蹇冩埧鑲岀粏鑳炵敓闀胯嚦80%铻嶅悎鏃讹紝寮冨幓鍩瑰吇鍩猴紝鐢ㄦ俯鑲茬殑PBS鍐叉礂缁嗚優2娆★紝姣忔10min锛岀劧鍚庣敤4%澶氳仛鐢查啗鍦ㄥ娓╂潯浠朵笅鍥哄畾缁嗚優15min锛汓/span>

2銆丳BS鍐叉礂缁嗚優2娆★紝姣忔10min锛岀劧鍚庡湪4鈩冩潯浠朵笅锛岀敤0.1锛匱riton X-100閫忚啘15min锛汓/span>

3銆丳BS鍐叉礂缁嗚優2娆★紝姣忔10min锛岀劧鍚庡湪瀹ゆ俯鏉′欢涓嬶紝鐢?% BSA灏侀棴缁嗚優30min锛汓/span>

4銆佹寜1: 100鐨勬瘮渚嬬█閲娢?actin涓€鎶楋紙Abbioscience鍏徃锛夛紝鐒跺悗灏嗗叾鏀惧湪4鈩冨啺绠变腑瀛佃偛缁嗚優杩囧锛汓/span>

5銆丳BS鍐叉礂缁嗚優3娆★紝姣忔10min锛屾寜1锛?50鐨勬瘮渚嬬█閲婃姉伪-actin鐨勪簩鎶楋紝 37鈩冩潯浠朵笅鏀剧疆1h锛汓/span>

6銆佺敤PBS鍐叉礂3娆★紝姣忔10min锛屾渶鍚庡湪鍊掔疆鑽у厜鏄惧井闀滀笅瑙傚療鍥惧儚骞舵媿鐓с€侟/span>