The culture medium was removed, and after washing the cells twice with PBS, they were harvested for further analysis. The preparation of cell lysates and Western blotting were performed by the method of Sun et al. [42 (link)]. Antibodies used were p16 INK4A (F-12) (Santa Cruz Biotechnology, sc-1661), P53 mouse monoclonal antibody (Proteintech, 60283-2-Ig), P21 rabbit polyclonal antibody (Proteintech, 27296-1-AP), LC3B antibody (Cell Signaling, 2575S), LAMP2 (Abcam, ab13524), AMPK (Cell Signaling, D63G4), P-AMPK (Cell Signaling, 2535S), p70 S6 Kinase Antibody (Cell Signaling, 9202S), p-p70 S6 Kinase Antibody (Cell Signaling, 9234S), and GAPDH polyclonal antibody (Proteintech, 10494-1-AP).
P70 s6 kinase antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The P70 S6 Kinase Antibody is a specific antibody that recognizes the P70 S6 kinase protein. P70 S6 kinase is a serine/threonine protein kinase that plays a role in the regulation of cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
P ampk, by Cell Signaling Technology (1 mentions)
Lamp2, by Abcam (1 mentions)
Gapdh polyclonal antibody, by Proteintech (1 mentions)
P21 rabbit polyclonal antibody, by Proteintech (1 mentions)
Lc3b antibody, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Ki67 antibody, by Abcam (1 mentions)
Anti caspase 3 antibody, by Cell Signaling Technology (1 mentions)
E cadherin antibody, by Proteintech (1 mentions)
Vimentin antibody, by Proteintech (1 mentions)
P53 antibody, by Proteintech (1 mentions)
Pcna antibody, by Proteintech (1 mentions)
Mmp 9 antibody, by Proteintech (1 mentions)
Akt antibody, by Proteintech (1 mentions)
N cadherin antibody, by Proteintech (1 mentions)
Bcl 2 antibody, by Proteintech (1 mentions)
Peroxidase conjugated secondary antibodies, by CWBIO (1 mentions)
Pakt ser473 antibody, by Cell Signaling Technology (1 mentions)
Rapamycin, by Meilun (1 mentions)
Bax antibody, by Proteintech (1 mentions)
3 protocols using p70 s6 kinase antibody
1
Cell Lysate Preparation and Western Blotting
2
Rapamycin and 5-FU Synergistic Effects on Colorectal Cancer
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Rapamycin and 5-FU were obtained from meilunbio (Dalian, China). The anti Caspase3 antibody, Cleaved Caspase3 antibody, p-AKT (ser 473) antibody, p-AKT (Thr 308) antibody, p70 S6 Kinase antibody, and p-p70S6 Kinase (Thr 389) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). The ki-67 antibody was purchased from Abcam (Cambridge, UK). The BCL-2 antibody, Bax antibody, AKT antibody, p53 antibody, E-Cadherin antibody, N-Cadherin antibody, Vimentin antibody, MMP-9 antibody and PCNA antibody were purchased from Proteintech (Wuhan, China). The GAPDH rabbit polyclonal antibody was obtained from XianZhi Biotech (Hangzhou, China). The peroxidase-conjugated secondary antibodies were obtained from CWBIO (Taizhou, China).
The human CRC cell line HCT-116 and SW-480 cells were purchased from iCell Bioscience Inc (Shanghai, China). The cells were cultured in a DMEM medium containing 10% FBS in a 37 °C incubator containing 5% CO2.
Female athymic nude mice (6–8 weeks old) were purchased from Guangdong Medical Laboratory Animal Center. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals. The Laboratory Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University approved all experimental protocols.
The human CRC cell line HCT-116 and SW-480 cells were purchased from iCell Bioscience Inc (Shanghai, China). The cells were cultured in a DMEM medium containing 10% FBS in a 37 °C incubator containing 5% CO2.
Female athymic nude mice (6–8 weeks old) were purchased from Guangdong Medical Laboratory Animal Center. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals. The Laboratory Animal Ethics Committee of the Third Affiliated Hospital of Guangzhou Medical University approved all experimental protocols.
3
Phosphorylation Analysis of Muscle Proteins
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The gastrocnemius and tibialis anterior muscles of eight-week-old mice were isolated, minced, and then homogenized in an NP40 Buffer supplied with protease inhibitors and phosphatase inhibitors (Leagene Biotechnology, Beijing, China). These protein lysates were then quantified and analyzed by western blot with antibodies as follows: phospho-4E-BP1 antibody (#9451, Cell Signaling Technology, Danvers, MA, USA), phospho-p70 S6 kinase antibody (#9234, Cell Signaling Technology, Danvers, MA, USA), phospho-CDK Substrate Motif antibody (#9477, Cell Signaling Technology, Danvers, MA, USA), 4E-BP1 antibody (#9644, Cell Signaling Technology, Danvers, MA, USA), p70 S6 kinase antibody (#2708, Cell Signaling Technology, Danvers, MA, USA) and GAPDH antibody (#A19056, ABclonal Technology, Wuhan, China). The dilution ratio of these antibodies for detecting phosphorylation was 1:500, while that of all other antibodies was 1:1000. The signal was detected in a chemiluminescence ChemiScope 4300Pro system and quantified with Clinx Image Analysis software.
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