As previously described (27 (link)), sections were deparaffinized, rehydrated, soaked in an antigen retrieval solution with citrate buffer, and incubated overnight at 4°C with a rabbit polyclonal antibody against p-ERK (9101; 1:200 dilution; Cell Signaling Technology) or cyclin E1 (ab7959; 1:200 dilution; Abcam). Biotinylated secondary antibody (SA00004-1; 1:200 dilution; Proteintech Group, Inc.) was then added and followed by incubation with a streptavidin-peroxidase conjugate for 30 min at room temperature. Images were captured under a fluorescence microscope (Nikon Corporation).
Ab7959
Manufactured by Abcam
Ab7959 is a primary antibody that can be used to detect the expression of the protein of interest in various experimental applications. The core function of this product is to specifically bind to and enable the detection of the target protein.
Automatically generated - may contain errors
Lab products found in correlation
Sa00004 1, by Proteintech (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Ab1801, by Abcam (1 mentions)
Ab95281, by Abcam (1 mentions)
β actin 8h10d10, by Cell Signaling Technology (1 mentions)
Sc 397, by Santa Cruz Biotechnology (1 mentions)
Sc 20685, by Santa Cruz Biotechnology (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti mouse secondary antibody, by Abcam (1 mentions)
Sc 528, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit secondary antibody, by Abcam (1 mentions)
Bca kit, by Keygen Biotech (1 mentions)
3 protocols using ab7959
1
Immunohistochemical Analysis of p-ERK and Cyclin E1
2
VSMC Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After cell treatment, VSMCs were washed with phosphate-buffered saline (PBS) and lysed in RIPA buffer (Biotech, Shanghai, China). After one freeze/thaw cycle, lysates were centrifuged. Protein concentration was determined by a BCA protein assay (Biotech, Shanghai, China) using bovine serum albumin as the standard. A quantity amounting to 10 μg of protein sample was subjected to SDS-polyacrylamide gel electrophoresis. Proteins were then transferred to an ECL nitrocellulose membrane (Millipore). Incubating the membrane in Superblock (Pierce) for 1 h blocked nonspecific binding. Membranes were then incubated overnight at 4°C in primary antibodies, PPARγ1, Cyclin D1, Cyclin B1/cdc2 and β-actin (AbCam: ab8924, ab95281, ab7959, ab1801). All primary antibodies dilution was 1:1000 in each reaction. The blots were washed three times with TBST buffer and then incubated for 1 h at room temperature with anti-rabbit secondary antibody conjugated with horseradish peroxidase. Western blot analysis was conducted according to standard procedures using Supersignal chemiluminescence detection substrate (Pierce).
3
Western Blot Analysis of Cell Cycle Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested and treated with lysis buffer on ice (KeyGen Biotech. Co., Ltd.), and a bicinchoninic acid assay (BCA) kit (KeyGen Biotech. Co., Ltd.) was used to quantify protein concentration. Equal amounts of protein were loaded in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels. After separation in the gel, the protein was transferred on a polyvinylidene fluoride (PVDF) membrane. Membranes were blocked in 2% bovine serum albumin (BSA) in Tris buffered saline with Tween 20 (TBS-T) for 1 h and then incubated overnight (4°C) with antibodies against CDC45 (sc-20685, 1:500; Santa Cruz Biotechnology), cyclin D1 (2978, 1:1000; Cell Signaling Technology), cyclin E1 (ab7959, 1:200; Abcam), CCNE2 (11935-1-AP, 1:500; Proteintech), p21 (sc-397, 1:500; Santa Cruz Biotechnology), p27 (sc-528, 1:200; Santa Cruz Biotechnology), or β-actin (8H10D10, 1:1000; Cell Signaling Technology). After being washed in TBS-T, membranes were incubated with goat antirabbit horseradish peroxidase (HRP)-conjugated secondary antibody (1:10,000; Abcam) or goat anti-mouse HRPconjugated secondary antibody (1:10,000; Abcam) for 2 h at room temperature. The blots were visualized by enhanced chemiluminescence (ECL) detection (Thermo Fisher Scientific). All experiments were repeated at least three times independently.
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!