Western blot analysis was performed to quantify the amount of target protein. The Western blot has been described previously.16 (link) HPV16 E6 (1:700, Abcam, Boston, MA, USA), HPV16 E7 (1:200, Bioss, Beijing, China), CAB39 (1:1000; Cell Signaling Technology, Beverly, MA, USA), PI3K(P85α) (1:1000; Proteintech, Wuhan, China), P-AKT(ser473) (1:1000; Proteintech), AKT (1:1000; Proteintech), GLUT1 (1:500; Wanleibio, China), and GAPDH (1:15000, Proteintech). After the membranes were further incubated with appropriate horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit IgG (1:5000; Proteintech) at 37°C for 2 h, the immunosignal was detected by using ECL Western blot kit (advansta, USA). The bands were analyzed with BioImaging systems (UVP Inc., Upland, CA, USA).
Ecl western blot kit
Manufactured by Advansta
Sourced in United States
The ECL Western blot kit is a laboratory equipment used for the detection and analysis of proteins in biological samples. It provides the necessary reagents and components to perform electrochemiluminescence (ECL)-based Western blotting, a widely used technique in molecular biology and biochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Glut1, by Wanlei (2 mentions)
Hpv16 e7, by Bioss Antibodies (2 mentions)
P akt ser473, by Proteintech (1 mentions)
Pi3k p85α, by Proteintech (1 mentions)
Hpv16e6, by Abcam (1 mentions)
Cab39, by Cell Signaling Technology (1 mentions)
Gapdh, by Proteintech (1 mentions)
Hif 1α, by Wanlei (1 mentions)
Hpv16 e6, by Bioss Antibodies (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Txnip, by Proteintech (1 mentions)
2 protocols using ecl western blot kit
1
Quantification of Protein Levels via Western Blot
2
Western Blot Assay for HPV16 E6/E7 and PTEN
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The Western-blot assays described in reference with PMID 3183982514. HPV16 E6 (1:200, Bioss Biotechnology Co., Ltd, Beijing, China), HPV16 E7 (1:200, Bioss Biotechnology), PTEN (total protein, 1:500, Wanleibio Co., Ltd, Shenyang, China), p-PTEN-S380 (serine 380 phosphorylated protein, 1:1000, Zenbio Co., Ltd., Chengdu, China), TXNIP (1:500; Proteintech Co., Ltd, Wuhan, China), HIF-1α (1:1000; Wanleibio), GLUT1 (1:500; Wanleibio), and GAPDH (1:1000, Cell Signaling Technology, Danvers, MA, USA). Membranes were further incubated with peroxidase-coupled anti-mouse or anti-rabbit IgG (1:5000; Proteintech) at 37°C for 2 h. Bound proteins were visualized using the ECL Western blot kit (advansta, USA), and their densities were measured using a BioImaging systems (UVP Inc., Upland, CA, USA). Protein bands were visualized using electrochemiluminescence substrate (Pierce) and detected by using BioImaging Systems (DNR, Jerusalem, Israel). GAPDH protein levels were used as the control group to calculate relative protein levels.
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