After determining the protein concentration, each AH sample (containing 4 mg of total AH protein) was mixed with 5× sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) loading buffer and denatured at 100°C for 5 minutes. Then, the samples were separated by 15% gradient acrylamide SDS-PAGE. The separated protein bands were transferred onto a polyvinylidene difluoride (PVDF) blotting membrane (Millipore, Burlington, MA, USA) and incubated with primary rabbit anti-human antibodies for IL-16R (1:1000; ab2573; Abcam, Cambridge, MA, USA), IL-6 (1:1000; ab151538; Abcam), and β-actin (1:1000; ab8226; Abcam) at 4°C overnight. The membranes were then incubated with a goat anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibody (1:1000; ab6721; Abcam) for 30 minutes at room temperature. The immunoblotted bands were visualized using an enhanced chemiluminescence reagent (Pierce ECL Western Blotting Substrate; Thermo Fisher Scientific). The gray band values of the blots were analyzed using Scion Image (Version: Beta 4.0.2 of Scion Image, Scion Corporation, Frederick, MD, USA).
Ab151538
Manufactured by Abcam
Sourced in United States
Ab151538 is a polyclonal antibody targeting the human CD3 epsilon protein. This antibody is suitable for use in various immunoassays and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab8226, by Abcam (1 mentions)
Scion image, by Techcomp Instruments (1 mentions)
Pvdf blotting membrane, by Merck Group (1 mentions)
Ab6721, by Abcam (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Ab8348, by Abcam (1 mentions)
Ab208113, by Abcam (1 mentions)
Ab33894, by Abcam (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
2 protocols using ab151538
1
Western Blot Analysis of AH Proteins
2
Endometrial Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The endometrial tissues of rats in each group were added with RIPA lysis buffer (Beyotime, Haimen, China). After homogenization in a glass homogenizer, centrifugation was conducted for 15 min at the rate of 12,000 rpm, and the supernatant was collected for SDS-PAGE electrophoresis. The isolated protein after electrophoresis was transfer to nitrocellulose membrane by electric transfer, closed at room temperature for 1 h in 5% skim milk-PBS solution, and incubated overnight at 4°C with primary antibodies MCP-1 (ab151538, Abcam), ICAM-1 (ab33894, Abcam), IL-6 (ab208113, Abcam), and TNF-α (ab8348, Abcam). After washing the membrane with PBS buffer for three times, secondary antibodies were added for 1 h of incubation at room temperature. Then, the membrane was washed for another three times with PBS buffer and developed by enhanced chemiluminescence. With GAPDH as the internal reference gene, the gray value ratio of target band to reference band was regarded as the relative expression level of proteins.
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!