Cells (1X106 cells) were lysed in RIPA buffer (Beyotime, Beijing, China), and the protein samples were segregated using SDS-PAGE. The samples were transferred onto nitrocellulose membranes, and the membranes were blocked in 5% skim milk. Membranes were incubated with primary antibodies: anti-WDR62 and anti-β-actin (1:2000), anti-p-DNA-PK (DNA-dependent protein kinase) and anti-Rad51 (1:2500), anti-p-ERK (extracellular signal-regulated kinase) and anti-ERK (1:3000), anti-p-JNK and anti-JNK (c-Jun N-terminal kinase) (1:3500), anti-p-p38 and anti-p38 (1:4000). The membranes were then incubated with secondary antibodies (1:4500), and subjected to chemiluminescence reagent kit (Beyotime) according to previous study [15 (link)]. All the antibodies were purchased from Abcam.
Chemiluminescence reagent kit
Manufactured by Beyotime
Sourced in China
The Chemiluminescence reagent kit is a laboratory equipment designed for the detection and quantification of specific biomolecules in various samples. The kit utilizes chemiluminescent reactions to generate light signals that can be measured and analyzed.
Automatically generated - may contain errors
Lab products found in correlation
7 protocols using chemiluminescence reagent kit
1
Western Blot Analysis of Cellular Signaling Pathways
2
Western Blot Analysis of NUP205 in Brain Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An appropriate amount of brain tissue was homogenized with RIPA lysate (EpiZyme, China) and a protease inhibitor (EpiZyme, China). The brain tissue was then split on ice for 30 min, and the protein supernatant was centrifuged at 12000 rpm for 30 min at 4 °C. The protein concentration was detected using a BCA kit (Biosharp, China). Briefly, the protein was boiled at 100 °C for 10 min in 4× loading buffer (Solarbio, China), separated by SDS-PAGE electrophoresis, and then transferred to a PVDF membrane (Bio-Rad, USA). After the membrane was sealed with 5% evaporated milk, NUP205 (1:1000; Proteintech, China) and β-actin (1:1000; Bioss, China) primary antibodies were added overnight at 4 °C. Then, the membrane was incubated in goat anti-rabbit IgG H&L antibody (1:2000; Bioss, China) at 37 °C for 1 h. Finally, the protein blots were developed with a chemiluminescence reagent kit (Beyotime Biotechnology), and ImagePro-Plus software (version 6.0) was used for the quantitative analysis.
3
Immunoblotting Analysis of Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues and cells were lysed to collect the supernatants. Protein samples were segregated and then transferred onto nitrocellulose membranes. Membranes were then incubated with primary antibodies: anti-TDAG51 and anti-GAPDH (1:2,000), anti-Bax (1:3,000), anti-cleaved caspase 3 and anti-cleaved PARP (1:4,000), and anti-SREBP-1 and anti-ANGPTL8 (1:5,000). The membranes were incubated with secondary antibodies (1:5,000) and then subjected to a chemiluminescence reagent kit (Beyotime). All antibodies were purchased from Abcam.
4
Western Blot Analysis of Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein of tissues and cells were segregated and then transferred onto a nitrocellulose membrane. Membrane was blocked and then incubated with primary antibodies: anti-CEMIP and anti-GAPDH (1:2000), anti-STAT3 and anti-p-STAT3 (1:2500), anti-AKT and anti-p-ANT (1:3000), anti-p65 and anti-p-p65 (1:3500). The membranes were then incubated with secondary antibodies (1:4000) and subjected to chemiluminescence reagent kit (Beyotime Biotechnology, Beijing, China). All the proteins were purchased from Abcam (Cambridge, MA, USA) according to previous research [15 ].
5
Cardiac Tissue Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cardiac tissues were lysed in RIPA buffer (Beyotime, Beijing, China). Protein samples were segregated using SDS-PAGE, and then transferred onto nitrocellulose membranes. Membranes were blocked in 5% dry milk, and then incubated with primary antibodies: anti-cleaved caspase-3 and anti-β-actin (1:2,000), anti-BAX and anti-BCL-2 (1:2,500), anti-collagen I and anti-collagen III (1:3,000), anti-p-JNK and anti-JNK (1:3,500), anti-p-p38 and anti-p38 (1:4,000), anti-TLR4 (1:4,500), anti-p-NF-κB and anti-NF-κB (1:5,000). The membranes were then incubated with secondary antibodies (1:6,000), and subjected to chemiluminescence reagent kit (Beyotime). All the proteins were purchased from Abcam (Cambridge, MA, USA).
6
Western Blot Analysis of Hippocampal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hippocampal tissues were lysed in radioimmunoprecipitation assay buffer with protease inhibitor cocktail (Beyotime, Beijing, China). Protein samples were segregated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and then transferred onto nitrocellulose membranes. Membranes were blocked in 5% dry milk, and then incubated with primary antibodies: anti-GATA4 and anti-GAPDH (1:2000), anti-BAX and anti-BCL-2 (1:3000), and anti-Sp1 and 6E10 (1:4000). The membranes were then incubated with secondary antibodies (1:5000), and subjected to chemiluminescence reagent kit (Beyotime). All the antibodies were purchased from Abcam (Cambridge, MA, USA).
7
CTRP6 Regulation of ERK Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Supernatant of lung tissues was segregated using SDS-PAGE and transferred onto nitrocellulose membranes. Membranes were blocked in 5% bovine serum albumin and incubated with primary antibodies: anti-CTRP6 and antiβ-actin (1:2000), anti-p-ERK and anti-ERK (1:3000). The membranes were then incubated with secondary antibodies (1:4000), and subjected to chemiluminescence reagent kit (Beyotime). All the proteins were purchased from Abcam.
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!