Team:KAIST-Korea/Notebook/Memo/Data
From 2010.igem.org
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<span style=font-size:15px> <b> pDRAW32 </b> </span><br> | <span style=font-size:15px> <b> pDRAW32 </b> </span><br> | ||
<b>Electrophoretic mobility shift assay (EMSA).</b><br> | <b>Electrophoretic mobility shift assay (EMSA).</b><br> | ||
- | The acute-phase response element (APRE; 5'-GCGCCTTCTGGGAAGATCCTTACGGGAATTCAG-3') double-stranded oligonucleotide probe for STAT3 binding was end-labelled using polynucleotide kinase and -32-ATP. The STAT3 EMSA was performed on equal amounts of whole total cell extracts from COS-1 cells prepared as described elsewhere. Binding reactions (20 l) contained 2 104 cpm of APRE probe in 10 mM Hepes at pH 7.8, 50 mM KCl, 1 mM EDTA, 5 mM MgCl2, 10% glycerol, 25 mM DTT and 2 g poly dI-dC. Competitive inhibition experiments were performed using a 20-fold molar excess of unlabeled oligonucleotides: WT (indicated above) and mutant (5'-GCGCCTTCTGGGCCTATCCTTACGGGCCGTCAG-3'). Supershift assay was performed by addition of antibodies to the Flag epitope. Binding reaction mixture was incubated at room temperature for 30 min. STAT3–APRE complexes were resolved on 5% non-denaturing polyacrylamide gels, and radiolabelled bands were visualized by autoradiography.<br> | + | The acute-phase response element (APRE; 5'-GCGCCTTCTGGGAAGATCCTTACGGGAATTCAG-3') double-stranded<br> oligonucleotide probe for STAT3 binding was end-labelled using polynucleotide kinase and -32-ATP. <br>The STAT3 EMSA was performed on equal amounts of whole total cell extracts from COS-1 cells prepared as described elsewhere.<br> Binding reactions (20 l) contained 2 104 cpm of APRE probe in 10 mM Hepes at pH 7.8, 50 mM KCl, 1 mM EDTA,<br>5 mM MgCl2, 10% glycerol, 25 mM DTT and 2 g poly dI-dC.<br> Competitive inhibition experiments were performed using a 20-fold molar excess of unlabeled oligonucleotides:<br> WT (indicated above) and mutant (5'-GCGCCTTCTGGGCCTATCCTTACGGGCCGTCAG-3'). Supershift assay was performed by addition of antibodies to the Flag epitope.<br> Binding reaction mixture was incubated at room temperature for 30 min. STAT3–APRE complexes were resolved on 5% non-denaturing polyacrylamide gels, and radiolabelled bands were visualized by autoradiography.<br> |
:http://www.nature.com/ncb/journal/v6/n6/full/ncb1138.html <br><br> | :http://www.nature.com/ncb/journal/v6/n6/full/ncb1138.html <br><br> | ||
</td> | </td> |
Revision as of 06:27, 4 August 2010
Data
NM_001024074.2 → NP_001019245.1
NM_001024075.1 → NP_001019246.1
NM_006895.2 → NP_008826.1
(3 kinds of isoform is obtained by alternative splicing) Human Coronaviral Spike protein.
Nucleotide sequence is in
protein sequence is in SARS starin nucleotide sequence is in
protein sequence is in
It is not easy to bring cDNA in here, so we should keep finding a better way. Sequence we used
2. JAK JAK.fasta 3. STAT STAT.fasta 4. GFP GFP.fasta ER, HER2 Receptor signal peptide sequence is P07110[1-24].
Anti-estrogen kappa/gamma chain M.musculus mRNA for anti-estrogen receptor IG JS34/32 kappa chain V region M.musculus mRNA for anti-estrogen receptor IG JS34/32 gamma chain V region
Mouse Anti-Estrogen Receptor alpha Monoclonal Antibody, Unconjugated, Clone 33 Mouse Anti-Estrogen Receptor beta Monoclonal Antibody, Unconjugated, Clone 14C8 Mouse Anti-Estrogen Receptor alpha Monoclonal Antibody, Unconjugated, Clone 6F11
Anti-estrogen Receptor The other antibody NCBI information pDRAW32 |