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).
Hpv16 e7
Manufactured by Bioss Antibodies
Sourced in China
The HPV16 E7 is a recombinant protein that represents the E7 oncoprotein of the human papillomavirus type 16 (HPV16). It is commonly used as a research tool in the study of HPV biology and the development of HPV-related diseases.
Automatically generated - may contain errors
Lab products found in correlation
Hpv16 e6, by Bioss Antibodies (4 mentions)
Glut1, by Wanlei (2 mentions)
Ecl western blot kit, by Advansta (2 mentions)
Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Cell Signaling Technology (2 mentions)
Gapdh, by Cell Signaling Technology (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)
Lamin b, by Wanlei (1 mentions)
Hif 1α, by Wanlei (1 mentions)
Txnip, by Proteintech (1 mentions)
Hif 2α, by Abcam (1 mentions)
Vascular endothelial growth factor (vegf), by Wanlei (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
5 protocols using hpv16 e7
1
Quantification of Protein Levels via Western Blot
2
Immunoblotting Assay for HPV16 Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The assays were performed as previously described.6 (link) Information about primary antibodies is as follows: HPV16 E6 (1:200,
Bioss Biotechnology Co., Ltd., Beijing, China), HPV16 E7 (1:200, Bioss
Biotechnology Co., Ltd., Beijing, China), LKB1 (1:1000, Cell Signaling
Technology, Danvers, MA, USA), Sp1 (1:1000, Cell Signaling Technology, Danvers,
MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000, Cell
Signaling Technology, Danvers, MA, USA).
Bioss Biotechnology Co., Ltd., Beijing, China), HPV16 E7 (1:200, Bioss
Biotechnology Co., Ltd., Beijing, China), LKB1 (1:1000, Cell Signaling
Technology, Danvers, MA, USA), Sp1 (1:1000, Cell Signaling Technology, Danvers,
MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000, Cell
Signaling Technology, Danvers, MA, USA).
3
HPV16 E6/E7 and LKB1 Signaling Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The assays were performed as previously described.9 Information about primary antibodies is as follows: HPV16 E6 (1:100, Bioss Biotechnology Co., Ltd., Beijing, China), HPV16 E7 (1:100, Bioss Biotechnology Co., Ltd., Beijing, China), KIF7 (1:1000, Proteintech, Wuhan, China), LKB1 (1:800, Proteintech, Wuhan, China), p‐LKB1 (Phospho‐Ser428) (1:800, Sangon Biotech, Shanghai, China), GAPDH (1:1000, Cell Signaling Technology, Danvers, MA, USA), and Lamin B (1:800, WanLeibio, Shenyang, China).
4
Western Blot Assay for HPV16 E6/E7 and PTEN
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
5
Western Blot Analysis of Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in the lysis buffer to obtain the total protein. After the concentration of the protein was measured by the Bradford method, 60 µg of protein was separated on a 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel and then transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford, MA, USA). The membrane was blocked with 5% skimmed milk and incubated with the following primary antibodies separately overnight at 4°C: HPV16 E6 (1:100; Bioss, Beijing, China), HPV16 E7 (1:100; Bioss, Beiing, China), LKB1 (1:1000; Cell Signaling Technology, Boston, MA, USA), HIF-2α (1:600; Abcam, Cambridge, MA, USA), VEGF (1:600; WanLei, China), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1000; Cell Signaling Technology, Boston, MA, USA).
After incubated with horseradish peroxidase (HRP)conjugated secondary antibody (anti-rabbit IgG), the membrane was developed in enhanced chemiluminescence (ECL). The density of each protein band was measured and normalized to the density of GAPDH by BioImaging System.
After incubated with horseradish peroxidase (HRP)conjugated secondary antibody (anti-rabbit IgG), the membrane was developed in enhanced chemiluminescence (ECL). The density of each protein band was measured and normalized to the density of GAPDH by BioImaging System.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!