Tissues were fixed overnight in Hartman’s fixative (Sigma) and processed for H&E staining and immunohistochemistry according to standard protocols (VanGompel and Xu, 2010 (link)). Immunostaining for PUM1, CDKN1B, phospho-Histone H3 were performed, following citrate buffer antigen retrieval, by incubation with anti-PUM1(1/50, Abcam), anti-CDKN1B (1/50, Abcam), and anti-phospho-H3 (1/50, CST) primary antibodies and detected using Biotin-Streptavidin HRP Detection Systems (ZSGB-BIO). For BrdU incorporation experiments, mice were injected intraperitoneally with 50 mg/kg BrdU in PBS and sacrificed 2 hr later, and tissue sections were analyzed by immunohistochemistry with anti-BrdU antibody (Invitrogen). TUNEL analysis was performed using the In Situ Cell Death Detection Kit from Roche according to the manufacturer’s instructions. A minimum of three randomly chosen discontinuous sections were used to determine positive cells in tubules.
Anti cdkn1b
Manufactured by Abcam
Sourced in United States
Anti-CDKN1B is a laboratory reagent used to detect and quantify the expression of the CDKN1B protein in biological samples. CDKN1B is a cyclin-dependent kinase inhibitor that plays a role in cell cycle regulation. This product can be used in various research applications, such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti brdu antibody, by Thermo Fisher Scientific (2 mentions)
Hartman s fixative, by Merck Group (2 mentions)
Anti pum1, by Abcam (2 mentions)
In situ cell death detection kit, by Roche (2 mentions)
Anti β actin, by Abcam (2 mentions)
Odyssey v2, by LI COR (1 mentions)
Anti satb2, by Abcam (1 mentions)
Anti runx3, by Abcam (1 mentions)
Odyssey imaging system, by LI COR (1 mentions)
Anti rabbit secondary antibody, by Abcam (1 mentions)
Anti active caspase 3, by Abcam (1 mentions)
Anti pro caspase 3, by Abcam (1 mentions)
Anti cyclin e1, by Abcam (1 mentions)
Anti cdk2, by Abcam (1 mentions)
Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions)
Bicinchoninic acid protein assay kit, by Beyotime (1 mentions)
Anti bax, by Abcam (1 mentions)
Anti atf4, by Abcam (1 mentions)
Anti cdkn1a, by Abcam (1 mentions)
Anti stat6, by Abcam (1 mentions)
4 protocols using anti cdkn1b
1
Histological Analysis of Cell Proliferation
2
Histological Analysis of Cell Proliferation
3
Western Blot Analysis of Cell Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was isolated from cell lysates with radio-immunoprecipitation assay buffer and quantified with a bicinchoninic acid protein assay kit (Beyotime Institute of Biotechnology). The proteins were resolved on 10% SDS and subsequently transferred to polyvinylidene difluoride membranes (Bio-Rad Laboratories, Inc.). Following blocking, the membranes were incubated with primary antibodies at 4°C overnight and subsequently incubated with an anti-rabbit secondary antibody (Abcam; 1:5,000) at room temperature for 1 h. The membranes were scanned on an Odyssey Imaging System and analyzed with Odyssey v2.0 software (LICOR Biosciences). The primary antibodies used in the present study were as follows: anti-CDKN1B (Abcam; 1:1,000), anti-SATB2 (Abcam; 1:1,000), anti-ATF4 (Abcam; 1:1,000), anti-Runx3 (Abcam; 1:1,000), anti-Cyclin E1 (Abcam; 1:1,000), anti-CDK2 (Abcam; 1:1,000), anti-Bax (Abcam; 1:1,000), anti-X-linked inhibitor of apoptosis protein (XIAP, Abcam; 1:1,000), anti-pro-caspase 3 (Abcam; 1:1,000), anti-active caspase 3 (Abcam; 1:1,000) and anti-β-actin (Abcam; 1:1,000). β-actin was used as an internal control.
4
Western Blot Analysis of Cell Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was carried out using anti‐STAT6 (1:2000), anti‐CDKN1A (1:1000), anti‐CDKN1B (1:1000), and anti‐β‐actin (1:1000) antibodies (all Abcam, Cambridge, MA, USA). The cells were detached with a cell lifter and lysed in RIPA lysis buffer (Solarbio Science and Technology). Approximately 25 μg protein was resolved on 12% T SDS–polyacrylamide gels and transferred to Immobilon‐P membranes (Millipore, Billerica, MA, USA). After blocking with 5% non‐fat milk, the membranes were incubated with primary antibodies at room temperature for 2 h, washed with Tris‐buffered saline containing 0.1% Tween‐20, and incubated with species‐compatible secondary antibodies conjugated to HRP (1:10 000). The signals were visualized by enhanced chemiluminescence (Amersham Life Sciences, Little Chalfont, UK), and the individual intensities of the protein bands were quantified with ImageJ software (National Institutes of Health, Bethesda, MD, USA).
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!