Materials
Name | Company | Catalog Number | Comments |
Dynabeads Protein A | Invitrogen | 10002D | |
Dynabeads Protein G | Invitrogen | 10004D | |
Anti-FLAG antibody | Sigma | F3165 | |
Anti-FLAG antibody | Sigma | F7425 | |
Anti-RHA antibody | Vaxron | PA-001 | |
TRizol LS reagent | Life technology | 10296-028 | |
RNase H | Ambion | AM2292 | |
Chloroform | Fisher Scientific | BP1145-1 | |
Isopropanol | Fisher Scientific | BP26184 | |
RNaeasy clean-up column | Qiagen | 74204 | |
Omniscript reverse transcriptase | Qiagen | 205113 | |
RNase Out | Invitrogen | 10777-019 | |
Protease inhibitor cocktail | Roche | 5056489001 | |
Triton X-100 | Sigma | X100 | |
NP-40 | Sigma | 98379 | |
Glycerol | Fisher Scientific | 17904 | |
Random hexamer primers | Invitrogen | N8080127 | |
Oligo-dT primers | Invitrogen | AM5730G | |
PCR primers | IDT | Gene specific primers for PCR amplification | |
Oligonucleotide for RNase H mediated cleavage | IDT | Anti-sense primer for target RNA | |
Trypsin | Gibco Life technology | 25300-054 | |
DMEM tissue culture medium | Gibco Life technology | 11965-092 | |
Fetal bovine serum | Gibco Life technology | 10082-147 | |
Tris base | Fisher Scientific | BP152-5 | |
Sodium chloride | Fisher Scientific | S642-212 | |
Magnesium chloride | Fisher Scientific | BP214 | |
DTT | Fisher Scientific | R0862 | |
Sucrose | Fisher Scientific | BP220-212 | |
Nitrocellulose membrane | Bio-Rad | 1620112 | |
Magnetic stand | 1.7 ml micro-centrifuge tube holding | ||
Laminar hood | For animal tissue culture | ||
CO2 incubator | For animal tissue culture | ||
Protein gel apparatus | Protein sample separation | ||
Protein transfer apparatus | Protein sample transfer | ||
Ready to use protein gels (4-15%) | Protein sample separation | ||
Table top centrifuge | Pellet down the sample | ||
Table top rotator | Mix the sample end to end | ||
Vortex | Mix the samples |
References
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