Reagent | Type | Manufacturer | Catalog # | Comments |
---|---|---|---|---|
D04 cells | Cells | Provided by Chris Schmidt, Queensland Institute of Medical Research (QIMR) Berghofer, Australia |
||
RPMI | Media | Life Technologies | 21875-034 | |
FBS | Reagent | Life Technologies | 10270106 | |
Effectene Transfection Reagent | Reagent | Qiagen | 301425 | Replaces Cell Line Nucleofector Kit V (10 RCT) Lonza VACA1003 |
Myc-BRAFWT vector | Plasmid | Shared by original author | ||
Myc-BRAFD594A vector | Plasmid | Shared by original author | ||
Empty vector | Plasmid | Shared by original author | ||
35 mm culture dishes | Materials | |||
PBS | Reagent | Gibco | 10010-023 | Original not specified |
Tris-HCl | Chemical | Specific brand information will be left up to the discretion of the replicating lab and recorded later |
||
NaCl | Chemical | |||
Igepal | Chemical | |||
Na3VO4 | Chemical | |||
NaF | Chemical | |||
Leupeptin | Chemical | |||
Rabbit α myc | Antibody | Abcam | ab9106 | |
Mouse α CRAF (for Western blotting) | Antibody | BD Transduction Laboratories | 610152 | |
Mouse α myc (9B11) (HRP conjugate) | Antibody | Cell Signaling Technology | 2040 | |
Protein G sepharose beads | Materials | Sigma | P3296 | |
NuPAGE Sample buffer | Buffer | Invitrogen | NP0007 | Original not specified |
SDS-Page gel (4–12%) | Western blot reagent | Invitrogen | NP0322BOX | Original not specified |
Nitrocellulose membrane (iBlot) | Western blot reagent | Invitrogen | IB301002 | Original not specified |
Ponceau stain | Western blot reagent | Sigma-Aldrich | P7170-1L | Original not specified |
Tris | Chemical | Sigma-Aldrich | T6066 | Original not specified |
Tween-20 | Chemical | Sigma-Aldrich | P1379 | Original not specified |
HPR-conjugated secondary antibody | Western blot reagent | Bio-Rad | 170-5047 | Original not specified |
ECL Detection Kit | Western blot reagent | Invitrogen | 32132 | Original not specified |