澧炲己鍨婤CA铔嬬櫧瀹氶噺璇曞墏鐩扅/span>

Enhanced Bicinchoninic Acid Kit for Protein Determination

璐у彿	缁勪唤A	缁勪唤B	缁勪唤C	BSA鏍囧噯鍝?/span>	瑙勬牸
PD-BCA-E2	63 ml	60 ml	3 ml	1 X 1 ml	鍙緵120涓瘯绠狘/span> 锛 鎴?鍧?6瀛旀澘娴嬭瘯鐢ㄣ€侟/span>
PD-BCA-E	250 ml	240 ml	12 ml	4 X 1 ml	鍙緵480涓瘯绠★紝 鎴?2鍧?6瀛旀澘娴嬭瘯鐢ㄣ€侟/span>

BSA鏍囧噯鍝佷负5 mg/ml BSA锛屾憾浜庢按涓紝鍚?.05%鍙燼zide姘挔銆侟/span>

瀹ゆ俯杩愯緭鍜屽偍瀛樸€傚鍥犳俯搴︿綆鎴栭暱鏃堕棿鏀剧疆鍑虹幇娌夋穩锛屽皢鍏舵憞鍖€鍗冲彲浣跨敤銆侭SA鏍囧噯鍝佷繚瀛樺湪-20搴︼紝濡傚嚭鐜板井鐢熺墿姹℃煋鍒欏簲涓㈠純銆侟/span>

浜у搧绠€浠婞/span>

BCA铔嬬櫧瀹氶噺鐨勫師鐞嗘槸鍩轰簬铔嬬櫧璐ㄤ笌閾滅瀛愬拰纰辨€ф煋鏂欙紙BCA锛夌殑鍖栧鍙嶅簲銆傚湪纰辨€ф潯浠朵笅锛岃泲鐧借川涓殑铔嬬櫧璐ㄩ叞鍩猴紙-NHCO-)涓嶣CA涓殑Cu2+绂诲瓙褰㈡垚绱壊缁滃悎鐗╋紝鍏跺惛鍏夊害涓庤泲鐧借川鐨勬祿搴︽垚姝ｆ瘮銆傚洜姝わ紝閫氳繃娴嬮噺鏍峰搧涓殑绱壊缁滃悎鐗╃殑鍚稿厜搴︼紝鍙互璁＄畻鍑烘牱鍝佷腑铔嬬櫧璐ㄧ殑娴撳害銆侟br/>

澧炲己鍨婤CA铔嬬櫧瀹氶噺璇曞墏鐩掗噰鐢ㄤ簡娴撶缉鍨嬭瘯鍓傞厤鏂癸紝鎻愰珮浜嗗鑲叉椂闂村拰娓╁害锛屽洜姝ゅ彲浠ュ鍔犳牱鍝佷綋绉拰澧炲姞鍏夊惛鏀跺€硷紝浠庤€岃兘澶熸娴嬫洿浣庢祿搴︾殑铔嬬櫧锛屾娴嬭寖鍥村彲浣庤嚦0.5鍒?0渭g/ml銆侟br/>

鏍囧噯铔嬬櫧鍜屽伐浣滄恫鐨勯厤鍒迭/span>

閰嶅埗鐗涜娓呯櫧铔嬬櫧姊害锛氬彲鏍规嵁妫€娴嬭寖鍥村拰棰勪及鐨勮泲鐧芥祿搴︽潵閰嶅埗BSA娴撳害姊害锛屽缓璁娇鐢ㄤ笌寰呮祴鏍峰搧鐩稿悓鐨勬憾鍓傘€傚彲鎸夋瘮渚嬫斁澶ф垨缂╁皬锛屾牴鎹疄闄呴渶瑕佽绠楁墍闇€鐢ㄩ噺銆侟/span>

BCA宸ヤ綔娑茬殑閰嶅埗

鏍规嵁闇€瑕佹祴瀹氱殑鏈煡鏍峰搧鍜岃泲鐧芥爣鍑嗗搧鐨勬暟閲忎互鍙婂瀛旀暟鏉ヨ绠楁墍闇€鐨凚CA宸ヤ綔娑蹭綋绉€傚浣跨敤璇曠杩涜娴嬭瘯锛屾瘡绠￠渶1.0 mL宸ヤ綔娑秉/span>,鑰屼娇鐢?6瀛旀澘杩涜娴嬭瘯鍒欐瘡瀛旈渶150 渭l宸ヤ綔娑层€侟/span>

鎸缁勪唤A:缁勪唤B:缁勪唤C = 25:24:1鐨勬瘮渚嬮厤鍒禕CA宸ヤ綔娑层€傜粍浠紺鍔犲叆缁勪唤A+B鐨勬椂鍊欙紝寮€濮嬩細鍑虹幇娴戞祳骞堕殢鐫€娣峰寑寰堝揩娑堝け锛屾渶缁堟贩鍖€鍚庢憾娑插憟鑻规灉缁胯壊銆傞厤缃ソ鐨勫伐浣滄恫鍦ㄥ娓╁瘑闂殑鏉′欢涓?4灏忔椂鍐呯ǔ瀹氥€侟/span>

鎿嶄綔姝ラ

濡傚湪璇曠涓繘琛屽弽搴旓紝鍙朁/span>1.0 ml鐨凚SA铔嬬櫧姊害鎴栬€呭緟娴嬫牱鍝侊紝鏀惧叆鏍囪濂界殑璇曠涓紝鍐嶆瘡绠″姞鍏?/span>1.0 ml宸ヤ綔娑诧紝鍏呭垎娣峰寑锛孅/span>鐩栦笂鐩栧瓙锛屽苟鍦?/span>60掳C瀛佃偛60minutes,妫€娴嬭寖鍥达細0.5-20 渭g/ml銆侟/span>

寤堕暱瀛佃偛鏃堕棿浼氬鍔犲厜鍚告敹鍊硷紱搴斾娇鐢ㄦ按娴存硶鍔犵儹锛屼互浣垮緱娓╁害鍧囪　涓婂崌锛涚敤榧撻娉曞崌娓╀細瀵艰嚧娓╁害涓嶅潎琛★紝浣垮疄楠岃宸澶с€侟/span>

濡傞渶鍦?6瀛旀澘涓繘琛屽弽搴旓紝鍒欐瘡瀛旀斁鍏?50 渭l鐨凚SA铔嬬櫧姊害鎴栬€呭緟娴嬫牱鍝侊紝鍐嶅姞鍏?50 渭l宸ヤ綔娑诧紝鍏呭垎娣峰寑锛?7搴﹀鑲?20鍒嗛挓锛屾娴嬭寖鍥翠负2-40 渭g/ml锛涙敞鎰忔俯搴︿笉瑕侀珮浜?7搴︼紝鍚﹀垯浼氬嚭鐜拌儗鏅崌楂樺拰鍙嶅簲寮傚父锛屽ぇ澶氭暟鑱氳嫰涔欑儻锛坧olystyrene锛夊井瀛旀澘浼氬湪60搴﹀嚭鐜伴浘鍖栫幇璞★紱搴斾娇鐢ㄦ按娴存硶鍔犵儹銆侟/span>

灏嗚瘯绠℃垨96瀛旀澘鍐峰嵈鑷冲娓┿€侟/span>

鐢ㄥ垎鍏夊厜搴﹁鎴栭叾鏍囦华鍦?62 nm澶勮鍙栧厜鍚告敹鍊? 鎵€鏈夋牱鍝佸簲鍦?0鍒嗛挓鍐呰瀹屻€傚洜涓築CA鍙嶅簲娌℃湁缁堢偣锛岄殢鐫€鏃堕棿鐨勬帹绉诲厜鍚告敹鍊间細缁х画涓婂崌锛屽湪10鍒嗛挓鍐呰瀹屾墍鏈夋牱鍝佸彲浠ラ伩鍏嶄骇鐢熸槑鏄剧殑璇樊銆侟/span>

灏咮SA铔嬬櫧鏍囧噯鍝佸拰寰呮祴鏍峰搧鐨勫厜鍚告敹鍊煎噺鍘荤┖鐧藉鐓у€硷紝浠ュ緱鍒扮殑铔嬬櫧鏍囧噯鍝佸厜鍚告敹鍊煎铔嬬櫧娴撳害浣滄爣鍑嗘洸绾匡紝鏍规嵁杩欎釜鏇茬嚎鏂圭▼璁＄畻寰呮祴鏍峰搧鐨勮泲鐧芥祿搴︺€侟/span>

娉ㄦ剰浜嬮」锛欬/span>

娴嬪畾铔嬬櫧娴撳害鏃讹紝寤鸿浣挎湭鐭ユ牱鍝佺殑铔嬬櫧娴撳害澶勪簬鏍囧噯鎷熷悎鏇茬嚎鐨勬帴杩戜腑闂翠綅缃紝杩欐牱鑾峰緱鐨勭粨鏋滄洿鍑嗙‘銆侟/span>

閫氳繃澧炲姞瀛佃偛鏃堕棿鎴栬€呮彁楂樻牱鍝佹瘮渚嬶紝鍙互鎻愰珮鍏夊惛鏀跺€硷紝浠庤€屾娴嬫洿浣庣殑铔嬬櫧閲忥紝浣嗘槸浼氱缉灏忔娴嬭寖鍥达紝鍙牴鎹疄闄呴渶瑕佽繘琛岃皟鏁达紝鍚屾椂娉ㄦ剰涓嶈瓒呰繃浠櫒鐨勬娴嬩笂闄愩€侟/span>

鏈€閫傛娴嬫尝闀夸负562 nm銆傚湪540 nm鍒?90 nm鍖洪棿涔熷彲杩涜妫€娴? 浣嗘爣鍑嗘洸绾挎枩鐜囧拰妫€娴嬬伒鏁忓害閮戒細鏈夋墍闄嶄綆銆侟/span>

閰舵爣浠殑妫€娴嬪厜寰勬瘮鍒嗗厜鍏夊害璁＄殑姣旇壊鏉煭锛屽洜鑰屾娴嬬伒鏁忓害鏈夋墍闄嶄綆銆侟/span>

鐢ㄨ蒋浠跺閰舵爣浠殑璇绘暟杩涜鏇茬嚎鎷熷悎鏃讹紝閲囩敤best-fit polynomial equation杩涜鎷熷悎鍙互鑾峰緱姣旂嚎鎬ф嫙鍚堟洿鍑嗙‘鐨勭粨鏋滐紱鎵嬪伐鎷熷悎鏃跺彲閲囩敤绾挎€ф嫙鍚堟硶銆侟/span>

EDTA娴撳害蹇呴』灏忎簬0.5 mM锛屼笉鍏煎EGTA銆備笉閫傜敤BCA娉曟椂锛岃浣跨敤Bradford铔嬬櫧娴撳害娴嬪畾娉曘€侟/span>

姣忕甯哥敤鐨勬€昏泲鐧芥祴瀹氭柟娉曟祴瀹氫笉鍚岀殑铔嬬櫧鏃堕兘鏈変竴浜涘亸宸€傚鑷翠骇鐢熻繖浜涘亸宸殑鍘熷洜鏈夛細铔嬬櫧璐ㄧ殑姘ㄥ熀閰稿簭鍒楋紝绛夌數鐐癸紝铔嬬櫧鐨勭粨鏋勶紝鐗瑰畾绉嶇被鐨勬皑鍩洪吀渚ч摼鍜岃緟鍩恒€傛湁浜涚绫荤殑姘ㄥ熀閰镐晶閾惧拰杈呭熀鑳藉鏄捐壊鍙嶅簲浜х敓寰堝ぇ鐨勫奖鍝嶃€傚ぇ閮ㄥ垎铔嬬櫧娴嬪畾鏂规硶閮戒娇鐢ㄧ墰琛€娓呯櫧铔嬬櫧锛圔SA锛夋垨鍏嶇柅鐞冭泲鐧?IgG)浣滀负鏍囧噯鍝佹潵鍒朵綔鏍囧噯鏇茬嚎銆傚鏋滈渶瑕佺壒鍒簿纭殑妫€娴嬬粨鏋滐紝鍙互浣跨敤寰呮祴铔嬬櫧鐨勭函鍝佹潵鍒朵綔鏍囧噯鏇茬嚎銆侟/span>

涓€浜涚墿璐ㄥ彲浠ュ共鎵癇CA鍙嶅簲锛屽杩樺師鍓傘€佺粶鍚堝墏锛屼互鍙婂己閰稿拰寮虹⒈銆傝繕鏈変竴浜涚墿璐ㄥ共鎵拌緝灏忥紝鍙涓嶈秴杩囨渶澶у吋瀹规祿搴﹀嵆涓嶅奖鍝嶅弽搴旂殑杩涜銆侟/span>

鏈骇鍝佷粎闄愪簬涓撲笟浜哄憳鐨勭瀛︾爺绌剁敤銆侟/span>

涓轰簡鎮ㄧ殑瀹夊叏鍜屽仴搴凤紝璇风┛瀹為獙鏈嶅苟鎴翠竴娆℃€ф墜濂楁搷浣溿€侟/span>