The largest database of trusted experimental protocols

2 protocols using anti cdk4

1

Immunohistochemical Examination of Cell Cycle Regulators

Check if the same lab product or an alternative is used in the 5 most similar protocols
The paraffin-embedded tissue sections were used for examination of HE staining. For the IHC staining, specimen’s tissue slides were de-paraffinized, rehydrated, and antigen retrieval. The tissue slides were blocked for at least 1 h at room temperature and then incubated with appropriate primary antibodies overnight at 4 ℃. The antibodies included anti-PRMT2 (1:100; Abcam), anti-CCNB1 (1:200; Santa Cruz Biotechnology), anti-CCND1 (1:100; Absin), anti-CDK4 (1:200; Wanleibio), and anti-H3R8me2a (1:500; Novus Biologicals). After careful washing, the slides were incubated with horseradish peroxidase (HRP) conjugates using DAB detection. For the IHC analysis, we quantitatively scored the tissue sections according to the percentage of positive cells and staining intensity. We assigned the following proportion scores: 1 if 0–25% of the tumor cells showed positive staining, 2 if 26–50% of cells were stained, 3 if 51–75% stained, and 4 if 76–100% stained.
+ Open protocol
+ Expand
2

Klotho Modulates Angiotensin II-Induced Vascular Remodeling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Dulbecco’s modified Eagle’s medium (DMEM) was purchased from HyClone (HyClone, Logan, CT, USA). Fetal bovine serum (FBS) was purchased from ABGENT (San Diego, CA, USA). We purchased penicillin, streptomycin, and Angiotensin II from Sigma-Aldrich (St. Louis, MO, USA). The Cell Counting Kit-8 (CCK8) was purchased from Dojindo (Kumamoto, Japan). The rat recombinant Klotho protein was from Cloud-Clone Corporation (Houston, TX, USA). The antibodies used in this study were as follows: mouse monoclonal PCNA (PC10), rabbit polyclonal NF-κB p65 (C-20), rabbit polyclonal p-NF-κB p65 (Ser 536), rabbit polyclonal ERK1/2 (K-23), rabbit polyclonal p-ERK1/2 (Thr202/Tyr204), rabbit polyclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (FL-335) (Santa Cruz Biotechnology, CA, USA); and rabbit polyclonal SM22α (Cloud-Clone Corp., Wuhan, China). Anti-CDK2, anti-CDK4, anti-cyclin D, and anti-cyclin E antibodies were purchased from Wanleibio, Shenyang, China, and anti-Akt and anti-phospho-Akt (Ser473) antibodies were from Cell Signaling (MA, USA). The reactivity with rat antigens is included in all of these antibodies. Through ImageJ software (NIH, USA), the signal intensity was quantified.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!