Warning: mysql_query() [function.mysql-query]: Unable to save result set in D:\wwwroot\hs21cn2043\wwwroot\includes\db.inc.php on line 50
Database error: Invalid SQL: select count(id) from pwn_comment where pid='518079' and iffb='1'
MySQL Error: 1194 (Table 'pwn_comment' is marked as crashed and should be repaired)
#0 dbbase_sql->halt(Invalid SQL: select count(id) from pwn_comment where pid='518079' and iffb='1') called at [D:\wwwroot\hs21cn2043\wwwroot\includes\db.inc.php:54] #1 dbbase_sql->query(select count(id) from {P}_comment where pid='518079' and iffb='1') called at [D:\wwwroot\hs21cn2043\wwwroot\comment\module\CommentContent.php:65] #2 CommentContent() called at [D:\wwwroot\hs21cn2043\wwwroot\includes\common.inc.php:551] #3 printpage() called at [D:\wwwroot\hs21cn2043\wwwroot\comment\html\index.php:13]
Warning: mysql_fetch_array(): supplied argument is not a valid MySQL result resource in D:\wwwroot\hs21cn2043\wwwroot\includes\db.inc.php on line 61

Warning: mysql_query() [function.mysql-query]: Unable to save result set in D:\wwwroot\hs21cn2043\wwwroot\includes\db.inc.php on line 50
Database error: Invalid SQL: select * from pwn_comment where pid='518079' and iffb='1' order by id limit 0,10
MySQL Error: 1194 (Table 'pwn_comment' is marked as crashed and should be repaired)
#0 dbbase_sql->halt(Invalid SQL: select * from pwn_comment where pid='518079' and iffb='1' order by id limit 0,10) called at [D:\wwwroot\hs21cn2043\wwwroot\includes\db.inc.php:54] #1 dbbase_sql->query(select * from {P}_comment where pid='518079' and iffb='1' order by id limit 0,10) called at [D:\wwwroot\hs21cn2043\wwwroot\comment\module\CommentContent.php:167] #2 CommentContent() called at [D:\wwwroot\hs21cn2043\wwwroot\includes\common.inc.php:551] #3 printpage() called at [D:\wwwroot\hs21cn2043\wwwroot\comment\html\index.php:13]
Warning: mysql_fetch_array(): supplied argument is not a valid MySQL result resource in D:\wwwroot\hs21cn2043\wwwroot\includes\db.inc.php on line 61
网友点评-L lysate and 70 l of 1 ?CPRG substrate (chlorophenol red-b-D-galactopyranoside) as outlined by-线缆测高仪,超声波测高仪, 手持式测高仪-上海交通大学科技园上海野豹企业发展公司
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发布于:2018-9-4 12:37:49  访问:83 次 回复: 篇
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L lysate and 70 l of 1 ?CPRG substrate (chlorophenol red-b-D-galactopyranoside) as outlined by
The control-conditioned medium (L1-CM) was prepared from a normal L1 cell line (ATCC CRL-2648) utilizing the exact same protocol as for the L Wnt3a cells.Luciferase gene reporter assayLRP5 LGK974 cost mutants and 10 FBS-DMEM medium CHO cells were plated at two ?104 PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/29069523 cells/well onto a 24-well plate and transfected 24 h later using Lipofectamine (Invitrogen). CHO cells had been plated at 1 ?105 cells/well on 6-well plates and transfected 24 h later applying Lipofectamine (Invitrogen) with 2 g of either WTLRP5, the mutant LRP5 construct or pcDNA3.1+. Four hours later 10 FBS-DMEM or PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/25580570 Wnt3a-CM or control L1CM was added to each nicely (see detailed description of CM production above), and cells had been collected 48 h immediately after transfection. Every experiment was performed in triplicate and repeated at the least 3 instances on separate occasions. Total RNA was isolated applying the E.Z.N.A. RNA isolation kit such as a treatment with RNase-free DNase (OMEGA Bio-Tek). RNA concentrations have been measured working with a NanoDropTM ND-1000 spectrophotometer (Thermo Scientific) and cDNA was synthesized by reverse transcript PCR (RT-PCR) from 1 g of extracted RNA applying the iScriptTM cDNA Synthesis kit (BioRad). Quantitative real-time PCR (qPCR) analyses of Tph1 and 5-Htr1b expression had been performed with especially created murine primers (out there on request), as well as the Tph1 and 5-Htr1b final results have been compared with those for any normal b-actin manage.L lysate and 70 l of 1 ?CPRG substrate (chlorophenol red-b-D-galactopyranoside) in accordance with the Stratagene guidelines. In both instances a VictorTM3 V 1420 Multilabel Counter (Perkin Elmer) was used, as well as the relative luciferase unit (RLU) was determined from the ratio in between the luciferase and b-galactosidase activities.Quantitative real-time polymerase chain reaction (qPCR)Chinese Hamster Ovarian (CHO) cells have been cultured with ten fetal bovine serum (FBS) (HyClone) in Dulbecco‘s modified Eagle‘s medium (DMEM) (BIOCHROM AG), plated on 10 cm plates and transfected with 3 g of WT-LRP5-mycHis or mutant construct working with Lipofectamine transfection reagent (Invitrogen) in line with the manufacturer‘s protocol. After 48 h of transfection, the cell medium was collected as well as the cells have been lysed using 1 Triton-X-homogenisation buffer. A 7.5 SDS-PAGE gel was prepared and 25 l of medium or cell lysate and 10 l of SDS-PAGE loading buffer had been loaded on the gel and analyzed under reducing circumstances. Western blot analysis was performed applying the Anti-Myc tag, clone 9E10 antibody (Upstate).Producing Wnt3a -conditioned mediaWnt3a-conditioned medium was made and collected from mouse L1 cells expressing Wnt3a (L Wnt3a; ATCC CRL-2647) as outlined by the manufacturer‘s instructions. The control-conditioned medium (L1-CM) was prepared from a standard L1 cell line (ATCC CRL-2648) employing precisely the same protocol as for the L Wnt3a cells.Luciferase gene reporter assayLRP5 mutants and ten FBS-DMEM medium CHO cells have been plated at two ?104 PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/29069523 cells/well onto a 24-well plate and transfected 24 h later employing Lipofectamine (Invitrogen).
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