5 × 106 HEK293T or HEK293 FRT were washed with ice-cold PBS and lysed with RIPA buffer containing protease and phosphatase inhibitors. For immunoblotting, 25 μg of protein were then fractionated on Tris–glycine SDS–PAGE gradient (4–15% acrylamide) gels (#3450123; Bio-Rad), transferred onto nitrocellulose membranes (Millipore), and detected with the indicated antibodies using an LI-COR detection system. Primary antibody used in this study: DYKDDDDK Tag Antibody (Flag, 2368; Cell Signalling Technology), A-Raf Antibody (4432; Cell Signalling Technology), LZTR1 (sc-390166; Santa Cruz Biotechnology), p44/42 MAPK (Erk1/2) (9107; Cell Signalling Technology), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (4370; Cell Signalling Technology), GAPDH (G8795; Sigma-Aldrich).
Dykddddk tag antibody flag
Manufactured by Cell Signaling Technology
The DYKDDDDK Tag Antibody (Flag) is a laboratory reagent used to detect and purify proteins tagged with the DYKDDDDK sequence, also known as the 'Flag' tag. This antibody specifically recognizes the Flag tag and can be used in various techniques, such as immunoprecipitation, Western blotting, and immunohistochemistry, to identify and isolate Flag-tagged proteins.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by Merck Group (2 mentions)
P44 42 mapk erk1 2, by Cell Signaling Technology (2 mentions)
Gapdh, by Merck Group (2 mentions)
Phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions)
Tris glycine sds page gradient 4 15 acrylamide gels, by Bio-Rad (1 mentions)
2 protocols using dykddddk tag antibody flag
1
Immunoblotting of HEK293 Cell Lysates
2
Western Blot Analysis of Protein Targets
Check if the same lab product or an alternative is used in the 5 most similar protocols
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5×10 6 HEK293T or HEK293 FRT were washed with ice-cold PBS and lysed with RIPA buffer containing protease and phosphatase inhibitors. For immunoblotting, 25 μg of protein was then fractionated on Tris-glycine SDS-PAGE gradient (4-15% acrylamide) gels (BioRad; #3450123), transferred onto nitrocellulose membranes (Millipore), and detected with the indicated antibodies using a LI-COR detection system. Primary antibody used in this study: DYKDDDDK Tag Antibody (Flag, Cell Signaling Technology, 2368), A-Raf Antibody (Cell Signaling Technology, 4432), LZTR1 (Santa Cruz Biotechnology, sc-390166), p44/42 MAPK (Erk1/2) (Cell Signaling Technology, 9107), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling Technology, 4370) GAPDH (Sigma-Aldrich, G8795).
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