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='546254' 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='546254' 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='546254' 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='546254' 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='546254' 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='546254' 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-LDRv3364-61c have been propagated in DH-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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LDRv3364-61c have been propagated in DH
Following 48 h, cell cultures had been lysed with 0.1 Triton X-100 and centrifuged at three,000 ?g for 15 min to eliminate bacterial pellet from the suspension. Pre-cleared cell lysate was SN-38 MedChemExpress incubated with His-tag primary agarose conjugate antibody, overnight at 4 . Samples have been resolved by electrophoresis on 12 Tris Cl gels, transferred to nitrocellulose membranes and blocked with five non-fat milk solution. Membrane was exposed to His principal antibody at a dilution of 1:1,000 (Santa PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/24652678 Cruz Biotechnology, Inc., Santa Cruz, CA, USA) for 1 h, followed by incubation with antirabbit IgG linked to AlexaFluor680 secondary antibody (1:5,000; SCH 58261 MedChemExpress Li-Cor, Lincoln, NE, USA) for 30 min. Reactivity was assessed with PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/21034305 Odyssey Imager (Li-Cor).CASPASE-1 ACTIVITY AND INHIBITION ASSAYSUninfected and infected macrophages with all the 20A11 mutant, M. tuberculosis H37Rv or 20A11 complemented (Rv3361c-65c) strains were assayed for in situ labeling and detection of active caspase-1, making use of the APO LOGIXTM-Carboxyfluorescein (FAM) kit (Bachem, Torrance, CA, USA). Staurosporine-treated U937 cells had been employed as a good control. Briefly, ten l of 30?Caspase-1 detection reagent (FAM VAD MK) had been added for the cells in 300 l medium of 96-well plate and incubated at 37 for two h. Wells have been gently washed two instances. Just after the final wash, 100 l of wash buffer had been added to experimental wells, and right away processed for fluorescence microscopy or fluorometric evaluation. Readings have been taken on a Cytofluorometer II (Biosearch, Bedford, MA, USA) with excitation at 488 nm and emission at 515?530 nm. A cell-permeable inhibitor I of caspase-1 at concentration of 10 M plus a caspase-inhibitor negative handle at a 10-Mwww.frontiersin.orgJanuary 2012 | Volume 2 | Report 281 |Danelishvili et al.Mycobacterium tuberculosis and macrophage apoptosisconcentration (irreversible cathepsin B inhibitor; CalBiochem, Los Angeles, CA, USA) have been utilized in caspase-1 inhibition assay. Macrophage monolayers in two-chamber slides had been treated with caspase-1 inhibitor or negative handle after which had been infected with all the 20A11 mutant strain. Apoptosis was quantified with TUNEL assay.IMMUNOPRECIPITATION AND IMMUNOBLOT ANALYSISLENTIVIRAL TRANSDUCTION FOR COLOCALIZATION OF M. tuberculosis AND HOST PROTEINSU937 cells, infected with M. tuberculosis wild-type and 20A11 mutant, have been scraped from tissue culture plates in 25 cm2 flasks right after 24 h, 2 or 4 days of infection, and lysed with 0.1 SDS.LDRv3364-61c have been propagated in DH10B E. coli, following
LDRv3364-61c had been propagated in DH10B E. coli, following the transformation into M. tuberculosis 20A11 mutant by electroporation. Transformants have been chosen on Middlebrook 7H10 agar plates containing apramycin 200 g/ml and screened by PCR for apramycin gene (50). We have constructed the -lactamase reporter vector for Mycobacterium and visualized bacterial protein translocation (secretion) inside the cytoplasm of macrophages. The assay uses technology-LACTAMASE (BlaC) ASSAY FOR PROTEIN SECRETIONThe pLDG13:His:Rv3364c construct was introduced into M. tuberculosis H37Rv and grown for 21 days, following which cells were harvested, disturbed inside a bead-beater, and processed for western blot to insure that His:Rv3364c protein was expressed in M.
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