The L02 cells and liver tissue samples were prepared as described above, and total protein was extracted. Western blotting was used to detect the target proteins of each sample, such as phospho-ERK1/2 (p-ERK1/2) (AM071, Beyotime), ERK1/2 (AF1051, Beyotime), phospho-p38 MAPK (Thr180) (p-p38) (AF5884, Beyotime), p38 MAPK (p38) (AF7668, Beyotime), phospho-JNK1/2 (Thr183/Tyr185) (p-JNK) (AF5860, Beyotime), JNK (AJ518, Beyotime), GAPDH (AF1186, Beyotime), phospho-AMPKα (Thr172) (p-AMPKα) (2535T, CST), AMPKα (t-AMPK) (2532S, CST), phospho-ACC (Ser79) (p-ACC) (3661S, CST), and ACC (t-ACC) (3676T, CST). The target protein bands of western blots were analyzed and quantified. The levels of p-AMPKα, p-ACC p-ERK1/2, p-p38 and p-JNK were normalized to those of AMPKα, ACC, ERK1/2, p38 and JNK, respectively, and are presented as the fold change compared to the control treatment.
Aj518
Manufactured by Beyotime
Sourced in China, United States
The AJ518 is a laboratory equipment designed for precision measurement. It features a digital display and a range of measurement capabilities. The core function of the AJ518 is to provide accurate and reliable data for scientific and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Af5860, by Beyotime (2 mentions)
Am071, by Beyotime (1 mentions)
Af1186, by Beyotime (1 mentions)
Af1051, by Beyotime (1 mentions)
Ab40772, by Abcam (1 mentions)
Ab157220, by Abcam (1 mentions)
Gtx128171, by GeneTex (1 mentions)
Af2000, by R&D Systems (1 mentions)
Gtx108181, by GeneTex (1 mentions)
Ag019, by Beyotime (1 mentions)
Ta 09, by ZSGB-BIO (1 mentions)
Mmp7 d4h5, by Cell Signaling Technology (1 mentions)
Gadph, by Abcam (1 mentions)
P38 mapk, by Beyotime (1 mentions)
P jnk, by Beyotime (1 mentions)
Cyclin d1, by Beyotime (1 mentions)
Ab 150591, by Abcam (1 mentions)
Mmp9 d6o3h, by Cell Signaling Technology (1 mentions)
4 protocols using aj518
1
Protein Expression Analysis in Cell and Tissue
2
Antibody Characterization for Cellular Pathways
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Antibodies were used as follows: anti-VPS35 (1:10,000, ab157220, Abcam, USA), anti-E-cadherin (1:10,000, ab40772, Abcam, USA), anti-N-cadherin (1:1000, 13116, Cell Signaling Technology, USA), anti-β-actin (1:2000, TA-09, ZSGB-BIO, China), anti-GAPDH (1:2000, AG019, Beyotime, China), anti-ROCK1 (1:500, WL01761, Wanleibio, China), anti-JNK (1:1000, GTX133806, Gene Tex, USA), anti-p-JNK (1:1000, AJ518, Beyotime, China), anti-Na/K ATPase (1:2000, 619736, ZEN BIO, China), anti-Frizzled 2 (1:1000, 24272-1-AP, Proteintech, USA), anti-ROR1 (1 μg/ml, AF2000, R&D system, USA), anti-MMP9 (1:1000, S1241, Bioworld, USA), p62 (1:1000, 5114, Cell Signaling Technology, USA), anti-Rac (1:500, WL02851, Wanleibio, China), anti-RhoA (1:500, WL02853, Wanleibio, China), anti-Rac (1:1000, ab76535, Abcam, USA), anti-RhoA (1:1000, bs-5330R, Bioss, USA), anti-p-p62 (1:1000, GTX128171, Gene Tex, USA), anti-FZD1 (1:1000, GTX108181, Gene Tex, USA).
3
Immunoblotting Analysis of Cell Signaling
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Immunoblotting analysis was performed as described.9 5% BSA and non‐fat milk were used as blocking reagent. The corresponding antibodies mainly included ZCCHC14 (ab‐150591, Abcam, USA), P42/44 (AM076, Beyotime, China), p‐P42/44 (AM071, Beyotime, China), cyclinD1 (AC853, Beyotime, China), MMP7 (D4H5; 3801, Cell Signaling, USA), MMP9 (D6O3H; 13 667, Cell Signaling, USA), GADPH (ab8245, Abcam, USA), JNK (AJ518; Beyotime, China), P‐JNK (AJ516, Beyotime, China), P38MAPK (AM065, Beyotime, China) and p‐P38MAPK (AM063, Beyotime, China),
4
Signaling Pathway Activation by LPH
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The cells were seeded on 6 cm dishes at a concentration of 2 × 105 cells per well and incubated in growth medium until confluency; then, the cells were serum-starved in α-MEM for 12 h. Then, 500 μg/mL of LPH was added to the culture medium before incubating for different time periods (0.5, 1, 2, 4, 8, 12, 24, and 36 h) to detect activation of the signaling pathway by Western blotting. Primary rabbit antibodies of extracellular signal regulated kinase 1/2 (ERK, 137F5, CST), phospho-ERK1/2 (Thr202/Try204, D13.14.4E, CST), p38 (AF1111, Beyotime), phospho-p38 (Thr180/Try182, AF5887, Beyotime), jun N-amino-terminal kinase (JNK, AJ518, Beyotime), phospho-JNK (Thr183/Try185, AF5860, Beyotime), Akt (C67E7, CST), phospho-Akt (Ser473, D9E, CST), and the loading control α/β-tubulin were used to examine the changes in MAPK and Akt signaling pathways.
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