The brain tissue of SN was separated after decapitation, as described in the section of ELISA Experiments (n = 5 for each group). Western blot analyses were conducted with the primary antibodies against mGluR5 (1 : 1000, Ab76316, Abcam), p-Akt (1 : 1000, 9271, CST), Akt (1 : 1000, 9272, CST), p-GSK-3β (1 : 1000, 9336, CST), GSK-3β (1 : 1000, Ab227208, Abcam), p-CREB (1 : 1000, 9198, CST), CREB (1 : 1000, 9197, CST), and β-Tubulin (1 : 500, Ab6046, Abcam), following the similar procedure reported previously [23 (link)].
Ab227208
Manufactured by Abcam
Sourced in United States
Ab227208 is a lab equipment product. It is a recombinant protein that can be used for various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab76316, by Abcam (1 mentions)
Ab6046, by Abcam (1 mentions)
Ab38449, by Abcam (1 mentions)
Protein extraction reagent, by Roche (1 mentions)
Ab216330, by Abcam (1 mentions)
Ab8805, by Abcam (1 mentions)
Ab181602, by Abcam (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
Ecl kit, by Thermo Fisher Scientific (1 mentions)
Bca protein kit, by Thermo Fisher Scientific (1 mentions)
Ab75745, by Abcam (1 mentions)
Ab137747, by Abcam (1 mentions)
Rpn2232, by GE Healthcare (1 mentions)
Ab60946, by Abcam (1 mentions)
Gapdh primary antibody, by Sangon (1 mentions)
3 protocols using ab227208
1
Western Blot Analysis of Signaling Proteins
2
Protein Expression Analysis in Cells
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Protein was extracted from cells by a protein extraction reagent (Roche, Switzerland), and measured by a BCA protein kit (Pierce Biotechnology, USA). 40-μg protein for each sample was electrophoresed in 10-15% SDS gel, then transferred to nitrocellulose membranes. After blocked with 5% BSA solution at room temperature for 1 h, membranes were incubated with primary antibodies at a dilution of 1/1000 at 4 °C overnight, including E-cadherin (#3195, cell signal technology, USA), Vimentin (#5741, cell signal technology, USA), Snail (#3879, cell signal technology, USA), c-MET (ab216330, Abcam, USA), total AKT (ab38449, Abcam, USA), p-AKT (ab8805, Abcam, USA), GSK3β (ab227208, Abcam, USA), p-GSK3β (#9322, cell signal technology, USA), or GAPDH (ab181602, Abcam, USA). Then, followed with secondary antibody (1:5000) at 37°C for 1h. The signals were detected by ECL Kit (Pierce Biotechnology, USA), and were analyzed by Image pro plus software.
3
Quantitative Western Blot Analysis of Liver Proteins
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The liver tissue of mice was extracted with Ripa lysate. Liver tissues were harvested and stored at -80℃for western blot analyses. The cells were lysed with biyuntian Western IP lysate to extract total protein. The total protein of each cell lysate was quantified by BCA protein quantitative Kit (all gold). Draw standard curve. Calculate the concentration of protein sample to be determined.
Adjust the sample concentration to be consistent. The primary antibodies used in the experiment are as follows: InsR(1: 1000 dilution, ab137747, abcam), P-InsR(1: 1000 dilution, ab60946, abcam), GSK-3β(1: 500 dilution, ab227208, abcam), P-GSK-3β(1; 500 dilution, ab75745, abcam), GAPDH primary antibody(1: 1000 dilution, D190090-0100, Sangon Bio), secondary antibody Sangon Bio. Ect chemical developer (GE health, rpn2232) was used to detect and capture protein bands. The data obtained by Western blotting were analyzed by image Proplus software.
Adjust the sample concentration to be consistent. The primary antibodies used in the experiment are as follows: InsR(1: 1000 dilution, ab137747, abcam), P-InsR(1: 1000 dilution, ab60946, abcam), GSK-3β(1: 500 dilution, ab227208, abcam), P-GSK-3β(1; 500 dilution, ab75745, abcam), GAPDH primary antibody(1: 1000 dilution, D190090-0100, Sangon Bio), secondary antibody Sangon Bio. Ect chemical developer (GE health, rpn2232) was used to detect and capture protein bands. The data obtained by Western blotting were analyzed by image Proplus software.
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