Table 2 Antibody and primer specifications
Primary Antibodies | ||||
|---|---|---|---|---|
Epitope | Manufacturer | Catalog no. | Dilution | |
Akt | Cell Signaling | 2920 | 1:2,000 | |
p-Akt (T308) | Cell Signaling | 2965 | 1:2,000 | |
p-Akt (S473) | Cell Signaling | 4060 | 1:2,000 | |
mTOR | Cell Signaling | 2983 | 1:2,000 | |
p-mTOR (S2448) | Cell Signaling | 2971 | 1:2,000 | |
p-mTOR (S2481) | Cell Signaling | 2974 | 1:2,000 | |
S6K | Cell Signaling | 9202 | 1:2,000 | |
p-S6K (T389) | Cell Signaling | 9206 | 1:2,000 | |
GSK3β | Abcam | Ab31826 | 1:2,000 | |
P-GSK3β (S9) | Cell Signaling | 9336 | 1:2,000 | |
4E-BP1 | Santa Cruz | Sc-81149 | 1:200 | |
p-4E-BP1 (T37/46) | Cell Signaling | 2855 | 1:2,000 | |
MURF1 | Abcam | Ab4125 | 1:2,000 | |
Secondary antibodies | ||||
Epitope | Fluorophore conjugate | Manufacturer | Catalog no. | Dilution |
Mouse Ig | Alexa 680 | Invitrogen | A21057 | 1:10,000 |
Goat Ig | Alexa 680 | Invitrogen | A21084 | 1:10,000 |
Rabbit Ig | Dylight 800 | Pierce | 35571 | 1:10,000 |
PCR primers | ||||
Target | Primers | |||
FOXO1 | Sense | GCCCAACCAAAGCTTCCCACAC | ||
Antisense | TGGACTGCTTCTCTCAGTTCCTGCT | |||
FOXO3 | Sense | GCTGGGTGCCAGGCTGAAGG | ||
Antisense | TTGGCAAAGGGTTTTCTCTGTAGGT | |||
FOXO4 | Sense | GATGAGGGCGAGGGACTGGA | ||
Antisense | TCCACATCTGAAGCAGGGGACA | |||
MURF1 | Sense | TGGGGGAGCCACCTTCCTCT | ||
Antisense | ATGTTCTCAAAGCCCTGCTCTGTCT | |||
Atrogin-1 | Sense | TGTTACCCAAGGAAAGAGCAGTATGGA | ||
Antisense | ACGGAGCAGCTCTCTGGGTTATTG | |||
HADHA | Sense | GCGAGTCTGAAGCTGCCTCCTAA | ||
Antisense | GGCACATGACTGCCTCATTCACA | |||
S26 | Sense | AACACCCCCACCCCGATTTAGAC | ||
Antisense | GAACTCAGCTCCTTACATGGGCTTT | |||
Cyclophillin | Sense | TGCAGACAAGGTCCCAAAGACAG | ||
Antisense | TGAAAGCAGGAACCCTTATAACCA | |||
β2-microglobulin | Sense | GCTGTGCTCGCGCTACTCTCTCT | ||
Antisense | TCTGCTGGATGACGTGAGTAAACCT | |||
GAPDH | Sense | CCTCCTGCACCACCAACTGCTT | ||
Antisense | GAGGGGCCATCCACAGTCTTCT | |||