鎮ㄦ墍鍦ㄤ綅缃細棣栭〉/浜у搧涓績/浜у搧璇︽儏

Phos-tag鈩 棰勫埗鑳?0渭mol/l锛?%锛?3瀛擖/h4>

铔嬬櫧纾烽吀鍖栫爺绌剁殑棰勫埗鑳迭img src="http://labchem.fujifilm-wako.com.cn/upfiles/images/201701/2214850785628047.jpg" title="2214850785628047.jpg" alt="鏂板搧閫熼€掑浘wako.jpg" style="margin: 0px; padding: 0px; font-family: Arial, 寰蒋闆呴粦, sans-serif; border: none; float: right;"/>

SuperSep Phos-tag鈩↑/span>



銆€銆€SuperSep Phos-tag鈩 鏄爺绌惰泲鐧界7閰稿寲鐨勬柟娉曪紝鏃犻渶鐗瑰紓鎬х7閰稿寲鎶椾綋鎴栬€呭悓浣嶇礌鏍囪銆侟/span>

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鈼咟/span>SuperSep Phos-tag鈩↑/span>


銆€銆€Phos-tag鈩 鏄竴绉嶉鍒惰兌锛岄鍏堝姞鍏ヤ簡 50 渭mol/L 鐨 Phostag鈩 Acrylamide锛屾墦寮€鍖呰鍗冲彲鐩存帴浣跨敤銆傞鍒惰兌涓惈鏈夐攲浣滀负閲戝睘绂诲瓙锛屽湪涓績鍑濊兌缂撳啿娑蹭腑淇濆瓨绋冲畾鎬у緢濂斤紝寰楀埌鐨勫甫鏉$粨鏋滀篃寰堟暣榻愩€侟/span>

銆€銆€纾烽吀鍖栬泲鐧藉拰闈炵7閰稿寲铔嬬櫧浣滀负涓嶅悓鏉″甫鍒嗙銆侟/span>

銆€銆€鍒嗙鍚庯紝鑳跺彲鐢ㄤ簬鑰冮┈鏂寒钃濇煋鑹诧紝鍏嶇柅鍗拌抗鍜岃川璋卞疄楠屻€侟/span>

宸插叕寮€鐨勯獙璇佽泲鐧藉垪琛紝璇风偣鍑狐/strong>




鈼咟/span>Phos-tag鈩 SDS-PAGE 鐨勫師鐞咟/span>

2.png

鈼咟/span>鍦 HighWire Search 涓婃悳绱㈠埌鐨勮鏂囨暟

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鈼咟/span>杩愮敤


鍒╃敤 p35 鐨勪笝姘ㄩ吀绐佸彉浣撶‘瀹 Cdk5 婵€娲 p35 鐨勭7閰稿寲浣嶇偣


銆€銆€p35 甯歌鐨勭7閰稿寲浣嶇偣鏄 Ser8 鍜 Thr138銆備絾鏄 Ser8 鍜 Thr138 浣嶇偣寰€寰€浼氬彂鐢熶笝姘ㄩ吀绐佸彉锛屼骇鐢燂紦绉嶇獊鍙樹綋锛圫er8 绐佸彉浣擄細S8A锛孴hr138 绐佸彉浣擄細T138A锛孲er8 鍜 Thr138 鍙岀獊鍙樹綋锛?A锛夈€傝繖锛撶绐佸彉浣撱€侀噹鐢熷瀷 p35銆丆dk5 鍜屾病鏈夋縺閰舵椿鎬х殑 Cdk5 閮芥潵婧愪簬 COS-7 缁嗚優銆傝繖浜涚粏鑳炶瑙f恫鐢?/span>Phos-tag鈩 SDS-PAGE 鍜 Western blotting 杩涜妫€娴嬶紙妫€娴嬫姉浣擄細p35 鎶椾綋锛夈€侟/span>

4.png

100 渭M Phos-tag 鈩 涓欑儻閰拌兒, 7.5% 鑱氫笝鐑叞鑳哄嚌鑳迭/span>

鍙槑纭7閰稿寲浣嶇偣鍜屾潯甯﹁縼绉荤巼鐨勫叧绯伙紒


- 娉抽亾锛戯紙鏉″甫 L2 鍜 L4锛夊拰娉抽亾锛曪紙鏉″甫 M1锛夛細p35 鍦 Cdk5 鐨勪綔鐢ㄤ笅鍙戠敓浜嗙7閰稿寲锛汓/span>

- 娉抽亾锛戯紙鏉″甫 L2 鍜 L4锛夊拰娉抽亾锛擄紙鏉″甫 L2 鍜 L4锛夛細鍦ㄦ棤婵€閰舵椿鎬 Cdk5 鐨勪綔鐢ㄤ笅锛屽ぇ绾︽湁涓€鍗 p35 铔嬬櫧鍦 Thr138 浣嶇偣鍙戠敓纾烽吀鍖栵紝鍚屾牱鍦 138 浣嶅彂鐢熺獊鍙樼殑 p35 铔嬬櫧浜︽槸濡傛銆侟/span>

娉抽亾锛 锛堟潯甯 M1锛夊拰娉抽亾锛栵紙鏉″甫 L3 鍜 L4锛夛細Ser8 鍜 Thr138 鏄富瑕佺殑纾烽吀鍖栦綅鐐癸紱

- 娉抽亾锛曪紙鏉″甫 M1锛夈€佹吵閬擄紬锛堟潯甯1鍜孡2锛夊拰娉抽亾锛橈紙鏉″甫M2锛夛細鏉″甫 M1 鏄 Ser8 鍜孴 hr138 閮藉彂鐢熺7閰稿寲鐨勬潯甯︼紱

鏉″甫 M2 鏄彧鏈 Ser8 纾烽吀鍖栫殑鏉″甫锛涙潯甯 L1 鍜 L2 鏄彧鏈 Thr138 纾烽吀鍖栫殑鏉″甫銆侟/span>

鈥 鏉″甫 L1 鍜 L3 涓殑锛告槸涓嶇‘瀹氬摢涓綅鐐瑰彂鐢熺7閰稿寲鐨勬潯甯︼紱

鈥?nbsp;鏉″甫L4鏄潪纾烽吀鍖栫殑 p35 銆侟/span>


銆愬弬鑰冩枃鐚€慄/span>

鈻 Quantitative Measurement of in Vivo Phosphorylation States of Cdk5 Activator p35 by Phos-tag 鈩 SDS-PAGE. T. Hosokawa, T. Saito, A. Asada, K. Fukunaga, and S. Hisanaga,Mol. Cell. Proteomics, Jun 2010;9: 1133 - 1143.

銆愮粨鏋滄彁渚涖€慄/span>

鐞嗗寲瀛︾爺绌舵墍 鑴戠瀛︾患鍚堢爺绌朵腑蹇 鍥炶矾鍔熻兘鐮旂┒鏍稿績 璁板繂鍔熻兘鐮旂┒鍥㈤槦 缁嗗窛鏅烘案锛圖r. T. Hosokawa锛堻/span>

棣栭兘澶у涓滀含 鐞嗗伐瀛︾爺绌剁 鐢熷懡绉戝涓撲笟 绁炵粡鍒嗗瓙鍔熻兘鐮旂┒瀹 涔呮案鐪熷競锛圖r. S. Hisanaga锛堻/span>



妫€娴嬪惈鏈 Dnmt1 纾烽吀鍖栨縺閰剁殑鐗囨

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鎴戜滑鍙互纭畾鍦ㄧ墖娈典腑鍚湁鐩殑婵€閰讹紒


鈶?nbsp;閲囩敤浜插拰鑹茶氨娉曚粠榧犺剳鎻愬彇娑蹭腑绾寲 GST-Dnmt1锛?-290锛夌粨鍚堣泲鐧键/span>

鈶 浣跨敤 0.3 M 鍜 1 M NaCl 鐨 DNA 绾ょ淮绱犳煴娲楄劚寰楀埌鐩殑铔嬬櫧

鈶 GST-Dnmt1锛?-290锛変綔涓轰綋澶栨縺閰跺疄楠岀殑鍙嶅簲搴曠墿

鈶 Phos-tag 鈩 SDS-PAGE 鐢ㄤ簬Western blotting锛岀‘瀹氳縼绉绘潯甯︿腑姣忎釜鐗囨鐨勬縺閰舵椿鎬?/span>


銆愬弬鑰冩枃鐚€慄/span>

The DNA-binding activity of mouse DNA methyltransferase 1 is regulated by phosphorylation with casein kinase 1delta/epsilon. Y. Sugiyama, N. Hatano, N. Sueyoshi, I. Suetake, S. Tajima, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and I. Kameshita, Biochem. J., May 2010; 427(3): 489-97.

銆愮粨鏋滄彁渚涖€慄/span>

楂樼煡澶у 缁煎悎鐮旂┒涓績 鐢熷懡銆佸姛鑳界墿璐ㄩ儴闂 瀹為獙瀹炰範鏈哄櫒璁炬柦 鏉夊北搴峰锛圖r. Y. Sugiyama锛堻/span>

棣欏窛澶у 鍐滃閮 搴旂敤鐢熺墿绉戝绉 鍔ㄧ墿鍔熻兘鐢熷寲瀛︾爺绌跺 榫熶笅鍕囷紙Dr. I. Kameshita锛堻/span>



浜岀淮鐢垫吵涓殑搴旂敤锛氬垎鏋 hnRNP K 纾烽吀鍖栧紓鏋勪綋


銆€銆€灏忛紶宸ㄥ櫖缁嗚優 J774.1 缁 LPS 鍒烘縺鍚庯紝瑁傝В缁嗚優锛岀粡杩囧厤鐤矇娣€娉曞垎绂诲緱鍒 hnRNP K銆傚湪浜岀淮鐢垫吵涓紝涓€缁存槸 IPG 鑳讹紝浜岀淮鏄 Phos-tag 鈩 SDS-PAGE锛屽彲鍒嗙 hnRNP K 鐨勫紓鏋勪綋銆傚埄鐢ㄨ川璋变华锛屽彲浠ョ‘璁や笉鍚岀殑鐐逛唬琛ㄤ笉鍚岀殑浜氬瀷鎴栦慨楗拌泲鐧姐€侟/span>

6.png

鍚屼竴涓瓑鐢电偣鐨勪綅缃笂锛屼笉鍚屼綅鐐瑰彂鐢熺7閰稿寲閮藉彲浠ヨ鍖哄垎寮€鏉?/span>

锛堜緥: spots 6 vs. 8 and spots 4 vs. 7锛堻/span>


銆愬弬鑰冩枃鐚€慄/span>

鈻 Characterization of multiple alternative forms of heterogeneous nuclear ribonucleoprotein K by phosphate-affinity electrophoresis. Y. Kimura, K. Nagata, N Suzuki, R. Yokoyama, Y. Yamanaka, H. Kitamura, H. Hirano, and O. Ohara, Proteomics, Nov 2010; 10(21): 3884-95.

銆愮粨鏋滄彁渚涖€慄/span>

妯花甯傜珛澶у 鐢熷懡绾崇背绯荤粺绉戝鐮旂┒绉 鐢熺墿浣撹秴鍒嗗瓙绯荤粺绉戝涓撲笟 鏈ㄦ潙寮ョ敓锛圖r. Y. Kimura锛夈€佸钩閲庝箙锛圖r. H. Hirano锛堻/span>

鐞嗗寲瀛︾爺绌舵墍 RCAI 灏忓師鏀迭/span>



鈼咟/span>澶囨敞


鏍峰搧鍒跺锛欬/span>

Phos-tag SDS-PAGE 瀵逛簬铔嬬櫧鏍峰搧涓殑鏉傝川闈炲父鏁忔劅锛屽挨鍏舵槸铻悎鍓傦紝閽掗吀锛屾棤鏈虹洂锛岃〃闈㈡椿鎬у墏杩欑被鐗╄川銆侟/span>

寮虹儓寤鸿鍦 Phos-tag SDS-PAGE 涔嬪墠閫氳繃 TCA 娌夋穩鎴栨笚鏋愭硶闄嶄綆鏉傝川鍚噺銆侟/span>


杞啘鍓嶅鐞嗭細

鍙︿竴涓繀椤荤殑姝ラ鏄湪杞啘鍓嶏紝鐢 EDTA 鍘婚櫎鍑濊兌涓殑閲戝睘绂诲瓙锛圡n2+ 鎴栬€匷n2+锛夛紱璇ユ楠ゅ彲鎻愰珮铔嬬櫧鐨勮浆鑶滄晥鐜囥€侟/span>

鈼忋€€鍒嗗埆鍑嗗 10 mmol/L 鍚 EDTA 鍜屼笉鍚 EDTA 涓ょ 1x transfer buffer銆侟/span>

鈼忋€€灏嗗嚌鑳舵蹈娉″湪鍚 10 mmol/L EDTA 鐨 1x transfer buffer锛岃嚦灏 20 鍒嗛挓锛屾俯鍜屾憞鏅冦€傛洿鎹㈡柊缂撳啿娑诧紝閲嶅锛撴銆侟/span>

鈼忋€€灏嗗嚌鑳舵蹈娉″湪涓嶅惈 10 mmol/L EDTA 鐨 1x transfer buffer锛?0 鍒嗛挓锛屾俯鍜屾憞鏅冦€侟/span>

鈼忋€€杞啘鎿嶄綔*銆侟/span>

* 寤鸿鐢ㄦ箍娉曡浆鑶滐紝浠ユ彁楂樿浆鑶滄晥鐜囥€侟/span>

7.png

鈼咟/span>璐ㄩ噺鎺у埗


銆€銆€姣忎竴鎵 SuperSep Phos-tag鈩紝鍑哄巶鍓嶅潎鏍规嵁鍏朵骇鍝佽鏍艰繘琛屾祴璇曪紝浠ヤ繚璇佸彲鍒嗙纾烽吀鍖栧拰闈炵7閰稿寲铔嬬櫧锛屼互鍙婁粬浠殑鍒嗙鎴愰兘鍦ㄦ甯稿弬鏁板唴銆侟/span>



鈼嗕繚瀛樻俯搴?/strong>


2-10鈩傸/span>



鈼嗕骇鍝佷俊鎭?/span>


鐢ㄤ簬 Bio-Rad 浼箰鐢垫吵浠?/span>

璐у彿

鍝佸悕

鐢垫吵浠?/span>

瑙勬牸

198-17981

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 7.5%, 17well,

83脳100脳3.9mm

Mini-PROTEAN 

Tetra Cell

(Bio-Rad Laboratories, Inc.)

5 鍧桙/span>

195-17991

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 12.5%, 17well,

83脳100脳3.9mm

5 鍧桙/span>




鐢ㄤ簬 Life Technologies 鐢垫吵浠?/span>


璐у彿

鍝佸悕

鐢垫吵浠?/span>

瑙勬牸

192-18001

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 7.5%, 17well,

100脳100脳6.6mm

XCell SureLock 

Mini-Cell

(Life Technologies, Inc.)


5 鍧桙/span>

199-18011

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 12.5%, 17well,

100脳100脳6.6mm

5 鍧桙br style="margin: 0px; padding: 0px; font-family: Arial, 寰蒋闆呴粦, sans-serif;"/>



鐢ㄤ簬 Wako EasySeperator 鐢垫吵浠?/span>


浜у搧缂栧彿浜у搧鍑濊兌娴撳害瀛旀暟鍖呰
192-17401SuperSep鈩 Phos-tag鈩 (50 渭mol/L)
Phos-tag鈩 棰勫埗鑳?0 渭mol/L
6.0%13瀛擖/span>5鍧桙/span>
199-173916.0%17瀛擖/span>5鍧桙/span>
195-173717.5%13瀛擖/span>5鍧桙/span>
192-173817.5%17瀛擖/span>5鍧桙/span>
193-1671110.0%13瀛擖/span>5鍧桙/span>
190-1672110.0%17瀛擖/span>5鍧桙/span>
195-1639112.5%13瀛擖/span>5鍧桙/span>
193-1657112.5%17瀛擖/span>5鍧桙/span>
193-1669115.0%13瀛擖/span>5鍧桙/span>
196-1670115.0%17瀛擖/span>5鍧桙/span>
197-1685117.5%13瀛擖/span>5鍧桙/span>
194-1686117.5%17瀛擖/span>5鍧桙/span>



鈼嗙浉鍏充骇鍝?/strong>


浜у搧缂栧彿浜у搧鍚嶇О瑙勬牸
058-07681EasySeparator
Phos-tag棰勫埗鍑濊兌鐨勯厤濂楃數娉虫Ы 
1 set

鐢熷懡绉戝


鍖栧


鍒嗘瀽


浠櫒


鑰楁潗


浜у搧涓績 > 鐢熷懡绉戝 > 铔嬬櫧鐮旂┒ > 纾烽吀鍖栬泲鐧键/a>

SuperSep Phos-tag鈩 棰勫埗鑳迭br style="margin: 0px; padding: 0px;"/>SuperSep Phos-tag鈩↑/p>

铔嬬櫧纾烽吀鍖栫爺绌剁殑棰勫埗鑳迭img src="http://labchem.fujifilm-wako.com.cn/upfiles/images/201701/2214850785628047.jpg" title="2214850785628047.jpg" alt="鏂板搧閫熼€掑浘wako.jpg" style="margin: 0px; padding: 0px; font-family: Arial, 寰蒋闆呴粦, sans-serif; border: none; float: right;"/>

SuperSep Phos-tag鈩↑/span>



銆€銆€SuperSep Phos-tag鈩 鏄爺绌惰泲鐧界7閰稿寲鐨勬柟娉曪紝鏃犻渶鐗瑰紓鎬х7閰稿寲鎶椾綋鎴栬€呭悓浣嶇礌鏍囪銆侟/span>


192-17401


鈼咟/span>SuperSep Phos-tag鈩↑/span>


銆€銆€Phos-tag鈩 鏄竴绉嶉鍒惰兌锛岄鍏堝姞鍏ヤ簡 50 渭mol/L 鐨 Phostag鈩 Acrylamide锛屾墦寮€鍖呰鍗冲彲鐩存帴浣跨敤銆傞鍒惰兌涓惈鏈夐攲浣滀负閲戝睘绂诲瓙锛屽湪涓績鍑濊兌缂撳啿娑蹭腑淇濆瓨绋冲畾鎬у緢濂斤紝寰楀埌鐨勫甫鏉$粨鏋滀篃寰堟暣榻愩€侟/span>

銆€銆€纾烽吀鍖栬泲鐧藉拰闈炵7閰稿寲铔嬬櫧浣滀负涓嶅悓鏉″甫鍒嗙銆侟/span>

銆€銆€鍒嗙鍚庯紝鑳跺彲鐢ㄤ簬鑰冮┈鏂寒钃濇煋鑹诧紝鍏嶇柅鍗拌抗鍜岃川璋卞疄楠屻€侟/span>

宸插叕寮€鐨勯獙璇佽泲鐧藉垪琛紝璇风偣鍑狐/strong>




鈼咟/span>Phos-tag鈩 SDS-PAGE 鐨勫師鐞咟/span>


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鈼咟/span>鍦 HighWire Search 涓婃悳绱㈠埌鐨勮鏂囨暟


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鈼咟/span>杩愮敤


鍒╃敤 p35 鐨勪笝姘ㄩ吀绐佸彉浣撶‘瀹 Cdk5 婵€娲 p35 鐨勭7閰稿寲浣嶇偣


銆€銆€p35 甯歌鐨勭7閰稿寲浣嶇偣鏄 Ser8 鍜 Thr138銆備絾鏄 Ser8 鍜 Thr138 浣嶇偣寰€寰€浼氬彂鐢熶笝姘ㄩ吀绐佸彉锛屼骇鐢燂紦绉嶇獊鍙樹綋锛圫er8 绐佸彉浣擄細S8A锛孴hr138 绐佸彉浣擄細T138A锛孲er8 鍜 Thr138 鍙岀獊鍙樹綋锛?A锛夈€傝繖锛撶绐佸彉浣撱€侀噹鐢熷瀷 p35銆丆dk5 鍜屾病鏈夋縺閰舵椿鎬х殑 Cdk5 閮芥潵婧愪簬 COS-7 缁嗚優銆傝繖浜涚粏鑳炶瑙f恫鐢?/span>Phos-tag鈩 SDS-PAGE 鍜 Western blotting 杩涜妫€娴嬶紙妫€娴嬫姉浣擄細p35 鎶椾綋锛夈€侟/span>


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100 渭M Phos-tag 鈩 涓欑儻閰拌兒, 7.5% 鑱氫笝鐑叞鑳哄嚌鑳迭/span>

鍙槑纭7閰稿寲浣嶇偣鍜屾潯甯﹁縼绉荤巼鐨勫叧绯伙紒


- 娉抽亾锛戯紙鏉″甫 L2 鍜 L4锛夊拰娉抽亾锛曪紙鏉″甫 M1锛夛細p35 鍦 Cdk5 鐨勪綔鐢ㄤ笅鍙戠敓浜嗙7閰稿寲锛汓/span>

- 娉抽亾锛戯紙鏉″甫 L2 鍜 L4锛夊拰娉抽亾锛擄紙鏉″甫 L2 鍜 L4锛夛細鍦ㄦ棤婵€閰舵椿鎬 Cdk5 鐨勪綔鐢ㄤ笅锛屽ぇ绾︽湁涓€鍗 p35 铔嬬櫧鍦 Thr138 浣嶇偣鍙戠敓纾烽吀鍖栵紝鍚屾牱鍦 138 浣嶅彂鐢熺獊鍙樼殑 p35 铔嬬櫧浜︽槸濡傛銆侟/span>

娉抽亾锛 锛堟潯甯 M1锛夊拰娉抽亾锛栵紙鏉″甫 L3 鍜 L4锛夛細Ser8 鍜 Thr138 鏄富瑕佺殑纾烽吀鍖栦綅鐐癸紱

- 娉抽亾锛曪紙鏉″甫 M1锛夈€佹吵閬擄紬锛堟潯甯1鍜孡2锛夊拰娉抽亾锛橈紙鏉″甫M2锛夛細鏉″甫 M1 鏄 Ser8 鍜孴 hr138 閮藉彂鐢熺7閰稿寲鐨勬潯甯︼紱

鏉″甫 M2 鏄彧鏈 Ser8 纾烽吀鍖栫殑鏉″甫锛涙潯甯 L1 鍜 L2 鏄彧鏈 Thr138 纾烽吀鍖栫殑鏉″甫銆侟/span>

鈥 鏉″甫 L1 鍜 L3 涓殑锛告槸涓嶇‘瀹氬摢涓綅鐐瑰彂鐢熺7閰稿寲鐨勬潯甯︼紱

鈥?nbsp;鏉″甫L4鏄潪纾烽吀鍖栫殑 p35 銆侟/span>


銆愬弬鑰冩枃鐚€慄/span>

鈻 Quantitative Measurement of in Vivo Phosphorylation States of Cdk5 Activator p35 by Phos-tag 鈩 SDS-PAGE. T. Hosokawa, T. Saito, A. Asada, K. Fukunaga, and S. Hisanaga,Mol. Cell. Proteomics, Jun 2010;9: 1133 - 1143.

銆愮粨鏋滄彁渚涖€慄/span>

鐞嗗寲瀛︾爺绌舵墍 鑴戠瀛︾患鍚堢爺绌朵腑蹇 鍥炶矾鍔熻兘鐮旂┒鏍稿績 璁板繂鍔熻兘鐮旂┒鍥㈤槦 缁嗗窛鏅烘案锛圖r. T. Hosokawa锛堻/span>

棣栭兘澶у涓滀含 鐞嗗伐瀛︾爺绌剁 鐢熷懡绉戝涓撲笟 绁炵粡鍒嗗瓙鍔熻兘鐮旂┒瀹 涔呮案鐪熷競锛圖r. S. Hisanaga锛堻/span>



妫€娴嬪惈鏈 Dnmt1 纾烽吀鍖栨縺閰剁殑鐗囨


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鎴戜滑鍙互纭畾鍦ㄧ墖娈典腑鍚湁鐩殑婵€閰讹紒


鈶?nbsp;閲囩敤浜插拰鑹茶氨娉曚粠榧犺剳鎻愬彇娑蹭腑绾寲 GST-Dnmt1锛?-290锛夌粨鍚堣泲鐧键/span>

鈶 浣跨敤 0.3 M 鍜 1 M NaCl 鐨 DNA 绾ょ淮绱犳煴娲楄劚寰楀埌鐩殑铔嬬櫧

鈶 GST-Dnmt1锛?-290锛変綔涓轰綋澶栨縺閰跺疄楠岀殑鍙嶅簲搴曠墿

鈶 Phos-tag 鈩 SDS-PAGE 鐢ㄤ簬Western blotting锛岀‘瀹氳縼绉绘潯甯︿腑姣忎釜鐗囨鐨勬縺閰舵椿鎬?/span>


銆愬弬鑰冩枃鐚€慄/span>

The DNA-binding activity of mouse DNA methyltransferase 1 is regulated by phosphorylation with casein kinase 1delta/epsilon. Y. Sugiyama, N. Hatano, N. Sueyoshi, I. Suetake, S. Tajima, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and I. Kameshita, Biochem. J., May 2010; 427(3): 489-97.

銆愮粨鏋滄彁渚涖€慄/span>

楂樼煡澶у 缁煎悎鐮旂┒涓績 鐢熷懡銆佸姛鑳界墿璐ㄩ儴闂 瀹為獙瀹炰範鏈哄櫒璁炬柦 鏉夊北搴峰锛圖r. Y. Sugiyama锛堻/span>

棣欏窛澶у 鍐滃閮 搴旂敤鐢熺墿绉戝绉 鍔ㄧ墿鍔熻兘鐢熷寲瀛︾爺绌跺 榫熶笅鍕囷紙Dr. I. Kameshita锛堻/span>



浜岀淮鐢垫吵涓殑搴旂敤锛氬垎鏋 hnRNP K 纾烽吀鍖栧紓鏋勪綋


銆€銆€灏忛紶宸ㄥ櫖缁嗚優 J774.1 缁 LPS 鍒烘縺鍚庯紝瑁傝В缁嗚優锛岀粡杩囧厤鐤矇娣€娉曞垎绂诲緱鍒 hnRNP K銆傚湪浜岀淮鐢垫吵涓紝涓€缁存槸 IPG 鑳讹紝浜岀淮鏄 Phos-tag 鈩 SDS-PAGE锛屽彲鍒嗙 hnRNP K 鐨勫紓鏋勪綋銆傚埄鐢ㄨ川璋变华锛屽彲浠ョ‘璁や笉鍚岀殑鐐逛唬琛ㄤ笉鍚岀殑浜氬瀷鎴栦慨楗拌泲鐧姐€侟/span>


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鍚屼竴涓瓑鐢电偣鐨勪綅缃笂锛屼笉鍚屼綅鐐瑰彂鐢熺7閰稿寲閮藉彲浠ヨ鍖哄垎寮€鏉?/span>

锛堜緥: spots 6 vs. 8 and spots 4 vs. 7锛堻/span>


銆愬弬鑰冩枃鐚€慄/span>

鈻 Characterization of multiple alternative forms of heterogeneous nuclear ribonucleoprotein K by phosphate-affinity electrophoresis. Y. Kimura, K. Nagata, N Suzuki, R. Yokoyama, Y. Yamanaka, H. Kitamura, H. Hirano, and O. Ohara, Proteomics, Nov 2010; 10(21): 3884-95.

銆愮粨鏋滄彁渚涖€慄/span>

妯花甯傜珛澶у 鐢熷懡绾崇背绯荤粺绉戝鐮旂┒绉 鐢熺墿浣撹秴鍒嗗瓙绯荤粺绉戝涓撲笟 鏈ㄦ潙寮ョ敓锛圖r. Y. Kimura锛夈€佸钩閲庝箙锛圖r. H. Hirano锛堻/span>

鐞嗗寲瀛︾爺绌舵墍 RCAI 灏忓師鏀迭/span>



鈼咟/span>澶囨敞


鏍峰搧鍒跺锛欬/span>

Phos-tag SDS-PAGE 瀵逛簬铔嬬櫧鏍峰搧涓殑鏉傝川闈炲父鏁忔劅锛屽挨鍏舵槸铻悎鍓傦紝閽掗吀锛屾棤鏈虹洂锛岃〃闈㈡椿鎬у墏杩欑被鐗╄川銆侟/span>

寮虹儓寤鸿鍦 Phos-tag SDS-PAGE 涔嬪墠閫氳繃 TCA 娌夋穩鎴栨笚鏋愭硶闄嶄綆鏉傝川鍚噺銆侟/span>


杞啘鍓嶅鐞嗭細

鍙︿竴涓繀椤荤殑姝ラ鏄湪杞啘鍓嶏紝鐢 EDTA 鍘婚櫎鍑濊兌涓殑閲戝睘绂诲瓙锛圡n2+ 鎴栬€匷n2+锛夛紱璇ユ楠ゅ彲鎻愰珮铔嬬櫧鐨勮浆鑶滄晥鐜囥€侟/span>

鈼忋€€鍒嗗埆鍑嗗 10 mmol/L 鍚 EDTA 鍜屼笉鍚 EDTA 涓ょ 1x transfer buffer銆侟/span>

鈼忋€€灏嗗嚌鑳舵蹈娉″湪鍚 10 mmol/L EDTA 鐨 1x transfer buffer锛岃嚦灏 20 鍒嗛挓锛屾俯鍜屾憞鏅冦€傛洿鎹㈡柊缂撳啿娑诧紝閲嶅锛撴銆侟/span>

鈼忋€€灏嗗嚌鑳舵蹈娉″湪涓嶅惈 10 mmol/L EDTA 鐨 1x transfer buffer锛?0 鍒嗛挓锛屾俯鍜屾憞鏅冦€侟/span>

鈼忋€€杞啘鎿嶄綔*銆侟/span>

* 寤鸿鐢ㄦ箍娉曡浆鑶滐紝浠ユ彁楂樿浆鑶滄晥鐜囥€侟/span>


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鈼咟/span>璐ㄩ噺鎺у埗


銆€銆€姣忎竴鎵 SuperSep Phos-tag鈩紝鍑哄巶鍓嶅潎鏍规嵁鍏朵骇鍝佽鏍艰繘琛屾祴璇曪紝浠ヤ繚璇佸彲鍒嗙纾烽吀鍖栧拰闈炵7閰稿寲铔嬬櫧锛屼互鍙婁粬浠殑鍒嗙鎴愰兘鍦ㄦ甯稿弬鏁板唴銆侟/span>



鈼嗕繚瀛樻俯搴?/strong>


2-10鈩傸/span>



鈼嗕骇鍝佷俊鎭?/span>


鐢ㄤ簬 Bio-Rad 浼箰鐢垫吵浠?/span>


璐у彿

鍝佸悕

鐢垫吵浠?/span>

瑙勬牸

198-17981

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 7.5%, 17well,

83脳100脳3.9mm

Mini-PROTEAN 

Tetra Cell

(Bio-Rad Laboratories, Inc.)

5 鍧桙/span>

195-17991

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 12.5%, 17well,

83脳100脳3.9mm

5 鍧桙/span>




鐢ㄤ簬 Life Technologies 鐢垫吵浠?/span>


璐у彿

鍝佸悕

鐢垫吵浠?/span>

瑙勬牸

192-18001

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 7.5%, 17well,

100脳100脳6.6mm

XCell SureLock 

Mini-Cell

(Life Technologies, Inc.)


5 鍧桙/span>

199-18011

SuperSep鈩 Phos-tag鈩 (50渭mol/L), 12.5%, 17well,

100脳100脳6.6mm

5 鍧桙br style="margin: 0px; padding: 0px; font-family: Arial, 寰蒋闆呴粦, sans-serif;"/>



鐢ㄤ簬 Wako EasySeperator 鐢垫吵浠?/span>


浜у搧缂栧彿浜у搧鍑濊兌娴撳害瀛旀暟鍖呰
192-17401SuperSep鈩 Phos-tag鈩 (50 渭mol/L)
Phos-tag鈩 棰勫埗鑳?0 渭mol/L
6.0%13瀛擖/span>5鍧桙/span>
199-173916.0%17瀛擖/span>5鍧桙/span>
195-173717.5%13瀛擖/span>5鍧桙/span>
192-173817.5%17瀛擖/span>5鍧桙/span>
193-1671110.0%13瀛擖/span>5鍧桙/span>
190-1672110.0%17瀛擖/span>5鍧桙/span>
195-1639112.5%13瀛擖/span>5鍧桙/span>
193-1657112.5%17瀛擖/span>5鍧桙/span>
193-1669115.0%13瀛擖/span>5鍧桙/span>
196-1670115.0%17瀛擖/span>5鍧桙/span>
197-1685117.5%13瀛擖/span>5鍧桙/span>
194-1686117.5%17瀛擖/span>5鍧桙/span>



鈼嗙浉鍏充骇鍝?/strong>


浜у搧缂栧彿浜у搧鍚嶇О瑙勬牸
058-07681EasySeparator
Phos-tag棰勫埗鍑濊兌鐨勯厤濂楃數娉虫Ы 
1 set



浜у搧缂栧彿浜у搧鍚嶇О浜у搧瑙勬牸浜у搧绛夌骇
193-16711SuperSep Phos-tag鈩?nbsp; (50 渭mol/L), 10%, 13 well 
Phos-tag 13瀛?0%棰勫埗鑳迭/td>
5鍧桙/td>-
190-16721SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 10%, 17 well 
Phos-tag 17瀛?0%棰勫埗鑳迭/td>
5鍧桙/td>-
195-16391SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 12.5%, 13 well 
Phos-tag 13瀛?2.5%棰勫埗鑳迭/td>
5鍧桙/td>-
193-16571SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 12.5%, 17 well 
Phos-tag 17瀛?2.5%棰勫埗鑳迭/td>
5鍧桙/td>-
193-16691SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 15%, 13 well 
Phos-tag 13瀛?5%棰勫埗鑳迭/td>
5鍧桙/td>-
196-16701SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 15%, 17 well 
Phos-tag 17瀛?5%棰勫埗鑳迭/td>
5鍧桙/td>-
197-16851SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 17.5%, 13 well 
Phos-tag 13瀛?7.5%棰勫埗鑳迭/td>
5鍧桙/td>-
194-16861SuperSep Phos-tag鈩?nbsp;(50 渭mol/L), 17.5%, 17 well 
Phos-tag 17瀛?7.5%棰勫埗鑳迭/td>
5鍧桙/td>-


鑱旂郴鍦板潃

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