The largest database of trusted experimental protocols

2 protocols using anti h3k27me1

1

Histone H3 Methylation Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
5 μg of histone was separated on a 12% SDS-PAGE for H3K27me3 and H3 detection. The blots were incubated with the primary antibody, mouse anti- H3K27me3 (1:800, Abcam, Cambridge, UK), anti-H3K27me2, anti-H3K27me1, anti- Suz12 monoclonal antibody (1: 500, Cell Signaling Technology, USA), and mouse anti-H3 monoclonal antibody (1:1,500,000, Abcam, Cambridge, UK) overnight at 4 °C, and then incubated with secondary anti-mouse HRP conjugated antibody (1:5000, Santa, USA) for 1 h at room temperature. The blots were developed with SuperSignal West Pico Chemiluminescence Substrate (Thermo, USA) and quantitated on densitometer (Bio-Rad, Universal HoodII, USA) using Quantity One software.
+ Open protocol
+ Expand
2

Western Blot Analysis of Histone Modifications

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein samples were resolved by SDS-PAGE and transferred to nitrocellulose membranes. The membranes were blocked in 3% BSA buffer (10 mM Tris-HCl, pH7.4, 150 mM NaCl, 1 mM EDTA, 0.1% Tween-20 and 3% BSA). Primary antibodies used for western blotting were: anti-H3K9me1 (Cell Signaling, #14186), anti-H3K9me2 (Cell Signaling, #4658), anti-H3K9me3 (Wako Diagnostics/Chemicals, 309–34839), anti-H3K27me1 (Cell Signaling, #7693), anti-H3K27me2 (EMD Millipore, #04–821), anti-H3K27me3 (Abcam, ab6002), anti-total H3 (EMD Millipore, 06–755), anti-V5 (Abcam, ab27671), anti-phospho ERK1/2 (Cell Signaling, #4370), anti-ERK1/2 (Cell Signaling, #4695), anti-G9a (Cell Signaling, #3306), anti-GLP (Abcam, ab135487), anti-LC3B (Cell Signaling, #3868), anti-MITF (C5, in-house), anti-active β-catenin (Cell Signaling, #8814), β-catenin (Cell Signaling, #9587), anti-DKK1 (Santa Cruz, sc-374574), anti-β-actin (Santa Cruz, sc-47778), anti-α-tubilin (Sigma Aldrich, T9026), anti-Lamin A/C (Cell Signaling, #4777) and anti-Lamin B (Cell Signaling, #12586). Appropriate secondary antibodies were used in 5% skim milk/TBST buffer. Protein bands were visualized using Western lightning plus ECL (Perkin Elmer) and quantified using ImageJ software.
+ 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!