Whole-cell extracts were prepared as described previously (Liu et al. 2013 (link)). In brief, equivalent amounts of protein (50 μg) were separated by sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (10 %) and transferred to nitrocellulose membrane (Bio-Rad, Hercules, CA). The membranes were subsequently incubated with the following primary antibodies: GTSE1 (Proteintech Group, Inc), AKT, phospho-AKT (Ser473), and cyclin B1 from CST (Cell Signaling Technology, Beverly, MA, USA), BCL-2 and Bax from Abcam (AbCam, Cambridge, MA, USA), and GAPDH (CST) was used as a loading control. The blots were visualized with a horseradish peroxidase-conjugated secondary antibody (Kangchen, Shanghai, China) and an enhanced chemiluminescence kit (Millipore, Bedford, MA).
Gtse1
Manufactured by Proteintech
Sourced in United States
GTSE1 is a protein that plays a role in the regulation of microtubule dynamics during cell division. It is involved in the control of chromosome segregation and spindle assembly. GTSE1 is expressed in a cell cycle-dependent manner, with its levels highest during mitosis.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Proteintech (2 mentions)
Enhanced chemiluminescence kit, by Merck Group (1 mentions)
Cyclin b1, by Cell Signaling Technology (1 mentions)
Bcl 2 and bax, by Abcam (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Gapdh, by Abcam (1 mentions)
Cyclin e1, by Proteintech (1 mentions)
Nanog, by Proteintech (1 mentions)
E cadherin, by Proteintech (1 mentions)
N cadherin, by Proteintech (1 mentions)
Vimentin, by Proteintech (1 mentions)
Desmoplakin, by Proteintech (1 mentions)
Cyclin d1, by Proteintech (1 mentions)
Itga2, by Abcam (1 mentions)
Goat anti rabbit or goat anti mouse secondary antibody, by Proteintech (1 mentions)
4 protocols using gtse1
1
Western Blot Analysis of Cell Signaling
2
Comprehensive Western Blotting Protocol
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Western blotting was performed according to the standard protocol. The primary antibodies, including GTSE1(Proteintech, 21,319–1-AP), Ki67(Proteintech, 27,309–1-AP),mutant p53(Abcam, ab32049), p53(Proteintech, 10,442–1-AP),Cyclin D1(Proteintech, 60,186–1-Ig),Cyclin E1(Proteintech, 11,554–1-AP), p21(Proteintech,60,214–1-Ig), E-Cadherin (Proteintech, 20,874–1-AP), Desmoplakin (Proteintech,25,318–1-AP), N-Cadherin (Proteintech,22,018–1-AP), Vimentin (Proteintech, 10,366–1-AP), Nanog (Proteintech, 14,295–1-AP), ABCG2(CST, 42078S), TAZ (CST, 4883S), YAP (CST, 14074S), ERK1/2(Affinity, AF0155), phospho-ERK1/2(Affinity, AF1015), AKT (CST, 2938S), Phospho-Akt (Thr308) (CST, 13038S), Phospho-Akt (Ser473) (CST, X4060S), FoxC2(CST, 12974S),Slug (CST, 9585 T),Twist1(CST, 46702S),Snail (Proteintech, MG-3879 T) and GAPDH (Proteintech, 60,004–1-Ig) were used at a dilution of 1:1000.
3
Immunohistochemical Evaluation of GTSE1 and ITGA2
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Immunohistochemistry was carried out on paraffin‐embedded tissue samples using antibodies against GTSE1 (1:400; Proteintech, Chicago, IL, USA) and integrin subunit alpha 2 (ITGA2) (1:400; Abcam, Cambridge, UK), followed by a standard avidin‐biotin detection protocol using 3‐amino‐9 ethylcarbazole. Sample staining was independently scored as 0 (negative), 1, 2, or 3 (weak, intermediate, and strong, respectively) by 3 pathologists, based on both the proportion of positively stained tumor cells and their staining intensity. GTSE1 protein expression was classified as high (IHC score of 2 or 3) or low (IHC score of 0 or 1) expression.
4
Western Blot Analysis of Protein Expression
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Total protein of cells was extracted with a radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor under the condition of an ice bath. The extracted protein and loading buffer were added and transferred onto 0.22-mm methanol-excited polyvinylidene fluoride membranes after SDS-PAGE electrophoresis, and then membranes were blocked with 5% nonfat milk. Next, we incubated membranes with the following primary antibodies at 4°C overnight: TAF15 (1:1,000) (Cell Signaling Technology, USA), MTF1 (1:1,000) (Proteintech, USA), YY2 (1:200) (Santa Cruz Biotechnology, USA), GTSE1 (1:1,000) (Proteintech, USA), and GAPDH (1:10,000) (Proteintech, USA), followed by incubation with related horseradish peroxidase (HRP)-conjugated secondary antibodies, including goat anti-rabbit or goat anti-mouse secondary antibody (1:10,000) (Proteintech, USA) at room temperature for 1.5 h. The relative integrated density values (IDVs) were observed based on GAPDH as a control.
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