Proteins were extracted from cultured cells or animal tissues by cell extract buffer (50 mM Tris-HCl, pH 8.0, 4 M urea and 1% Triton X-100) with protease inhibitor mixture (Roche Diagnostics, Cat.No: 04693132001). Cell lysates were resolved by SDS-PAGE and then analyzed by western blotting. The following antibodies were used: ACTB (Proteintech, #20536-1-AP, 1:5000), PFKL (Cell Signaling Technology, #8175, 1:1000), PFKFB3 (Cell Signaling Technology, #13123, 1:1000), HK2 (Cell Signaling Technology, #2867, 1:1000), pyruvate dehydrogenase (Cell Signaling Technology, #3205, 1:1000), SDHA (Cell Signaling Technology, #11998, 1:1000), VDAC (Cell Signaling Technology, #4661, 1:1000), ATF4 (Cell Signaling Technology, #11815, 1:1000), P-eIF2α (Cell Signaling Technology, #3398, 1:1000), α-tubulin (SIGMA-ALDRICH, T9026, 1:500), SLC38A5 (abcam, ab72717, 1:1000), TIGA1 (Santa Cruz, sc-244,315, 1:200), PGC-1a (Cell Signaling Technology, #2178, 1:1000), P53 (Millipore, 17-613, 1:1000), SN2 (abcam, ab72717, 1:1000), HIF-1a (BD Pharmingen, 565,924, 1:200), H3K27ac (abcam, ab4729,1:1000), Anti-Rabbit IgG (KPL, 074-1506, 1:5000), Anti-mouse IgG (KPL, 074-1806, 1:5000), Anti-Goat IgG (KPL, 14-13-06, 1:5000).
Slc38a5
Manufactured by Abcam
SLC38A5 is a sodium-coupled neutral amino acid transporter that mediates the uptake of neutral amino acids, such as asparagine, glutamine, histidine, and serine, into cells.
Automatically generated - may contain errors
Lab products found in correlation
α tubulin, by Merck Group (1 mentions)
Protease inhibitor mixture, by Roche (1 mentions)
Pfkfb3, by Cell Signaling Technology (1 mentions)
Pgc 1a, by Cell Signaling Technology (1 mentions)
P eif2α, by Cell Signaling Technology (1 mentions)
H3k27ac, by Abcam (1 mentions)
Hif1a, by BD (1 mentions)
E cadherin, by BD (1 mentions)
Srebp1, by Santa Cruz Biotechnology (1 mentions)
N cadherin, by BD (1 mentions)
Stat3, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti rabbit 7074s, by Cell Signaling Technology (1 mentions)
Phospho mtor, by Cell Signaling Technology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
2 protocols using slc38a5
1
Western Blot Analysis of Cellular Proteins
2
Western Blotting for Protein Analysis
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Western blotting was performed as previously described [22 (link)]. Cell lysates were separated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. After blocking with 5% skim milk for 1 h, the membranes were incubated with primary antibodies (1:1000) at 4 °C overnight, followed by incubation with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000) for 1 h. The protein bands were then exposed to an enhanced chemiluminescent HRP substrate (Waltham, MA, USA) and detected using X-ray film. The primary antibodies: GAPDH (# sc-365062), NRF2 (# sc-365949), GPX4 (# sc-166570), SREBP1 (# sc-17755), and SCD1 (# sc-515844) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). mTOR (# 2972S), Phospho-mTOR (# 5536S), PI3K (# 4292S), pAKT (# 9271S), AKT (# 9272S), pSTAT3 (# 9145S), and STAT3 (# 9139S) were purchased from Cell Signaling Technology (Danvers, MA, USA). SLC38A5 (# ab72717) and RRM1 (# ab226391) were purchased from Abcam. N-cadherin (# 610920) and E-cadherin (# 610181) antibodies were purchased from BD Biosciences (Franklin Lakes, NJ, USA). Horseradish peroxidase-conjugated goat anti-mouse (# 7076S) and HRP-conjugated goat anti-rabbit (#7074S) secondary antibodies were purchased from Cell Signaling Technology.
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