Whole-cell lysates were separated via 10% SDS-PAGE and transferred onto PVDF membranes (Bio-Rad, CA, USA). Blots were blocked overnight at 4 °C in 5% non-fat milk, followed by incubation at room temperature with appropriate antibodies for 2 h. Antibodies utilized included anti-LRRC1 (1:1000, ab127568, Abcam), anti-PPARγ (1:1500, ab272718, Abcam), anti-CEBP/β (1:1000, ab53138, Abcam), anti-FASN (1:1000, ab128870, Abcam), anti-LIPE (1:1000, ab45422, Abcam), anti-SCD1 (1:1000, ab236868, Abcam), and anti-β-actin (1:100, ab6276, Abcam). Secondary HRP-conjugated antibodies (1:10,000) were used to detect protein bands, which were then detected via enhanced chemiluminescence system and analyzed with the Image Lab software.
Ab53138
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab53138 is a primary antibody product from Abcam. It is a rabbit polyclonal antibody that recognizes a specific target protein. This antibody is intended for use in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab127568, by Abcam (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Ab128870, by Abcam (1 mentions)
Ab45422, by Abcam (1 mentions)
Ab236868, by Abcam (1 mentions)
Ab272718, by Abcam (1 mentions)
Ab6276, by Abcam (1 mentions)
Ecl plus reagent, by Thermo Fisher Scientific (1 mentions)
Sa00001 1, by Proteintech (1 mentions)
Sa00001 2, by Proteintech (1 mentions)
Bicinchoninic acid kit, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Ab205718, by Abcam (1 mentions)
Ab9485, by Abcam (1 mentions)
3 protocols using ab53138
1
Western Blot Analysis of Lipid Metabolism Proteins
2
Quantitative Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bronchial epithelial cells were treated with 200 μl RIPA for 10 min on ice, and then were centrifuged at 12,000×g (4 °C) for 15 min. The loaded proteins (50–170 μg) were separated on a 10% SDS-PAGE, followed by transferring onto PVDF membranes. The samples were blocked with TBS-Tween 20 (TBST) containing 5% skim milk for 60 min at room temperature, and at 4 °C overnight, then 30 min at room temperature. The membranes were incubated with rabbit anti-mouse antibody against mouse C-EBPβ (1:1000, ab53138; Abcam, Cambridge, UK), and mouse anti-mouse antibody against β-actin (1:5000, 60008-1-Ig; Proteintech, IL, USA) for 90 min at room temperature, and then they were incubated with horseradish peroxidase conjugated goat anti-rabbit (1:6000, SA00001-2; Proteintech, IL, USA) or anti-mouse antibody (1:5000, SA00001-1; Proteintech, IL, USA) for 90 min at room temperature. At last, blots were developed with the ECL Plus reagents (Thermo pierce, IL, USA).
3
miR-191 Regulates C/EBPβ Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 cells were transfected with miR-191 for 48 h, as detailed above. For the immunoblotting analysis of C/EBPβ, the cells were lysed in ice-cold lysis buffer containing a protease inhibitor cocktail (Roche Diagnostics, Basel, Switzerland), and the protein extracts were denatured in a boiling water bath for 10 min. The concentration of protein was determined using a bicinchoninic acid kit (Thermo Fisher Scientific, Inc.). Total protein (30 µg) was separated by 10% SDS-PAGE and transferred to nitrocellulose membranes. Following blocking with 5% skimmed milk overnight at 4°C, the membranes were incubated with primary antibodies against C/EBPβ (ab53138; 1:1,000) and GAPDH (ab9485; 1:1,000; both Abcam, Cambridge, MA, USA) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated secondary antibodies (ab205718; 1:2,000; Abcam) at room temperature for 60 min. The target protein was detected with a chemiluminescent kit (GE Healthcare Life Sciences, Little Chalfont, UK). Protein was quantitatively analyzed using Image J 1.48 u 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!