After boiling in Laemmli’s sample buffer, proteins were resolved on 7% or 12% SDS-polyacrylamide gels, and transferred to the PVDF membrane. Membranes were incubated with a primary antibody. GR was detected using an anti-GR antibody (PA1 511A) (Affinity BioReagents, Golden, CO, USA). SOD1, SOD2, CAT, GSH-Red, GSH-Px, GLUT2, and 11βHSD1 were detected using Abcam (Abcam, Cambridge, UK) antibodies (ab13498, ab13533, ab16731, ab16801, ab22604, ab54460, and ab393364, respectively). β-actin was detected by AC-15 antibody (Sigma-Aldrich, St. Louis, MO, USA). NF-κB-p65 subunit, lipin-1, and lamin B were detected using antibodies from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Dallas, TX, USA) (sc-372, sc-98450, and sc-6217, respectively). After being incubated with alkaline phosphatase linked secondary antibody (Amersham Pharmacia Biotech, Little Chalfont, UK), immunoreactive proteins were visualized by the enhanced chemifluorescence method (Amersham Biosciences). Quantitative analysis of immunoreactive bands was done using ImageQuant software. β-actin was used as equal load controls for whole cell extracts and cytosols, and lamin B for nucleosols.
Ab393364
Manufactured by Abcam
Sourced in United Kingdom
Ab393364 is a laboratory product manufactured by Abcam. It is a tool used for scientific research purposes. The core function of this product is to provide a specific type of laboratory equipment or reagent. No further details about the intended use or application of this product can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Ab13533, by Abcam (1 mentions)
Ab13498, by Abcam (1 mentions)
Ab16731, by Abcam (1 mentions)
Ab16801, by Abcam (1 mentions)
Alkaline phosphatase linked secondary antibody, by Cytiva (1 mentions)
Sc 6217, by Santa Cruz Biotechnology (1 mentions)
Sc 372, by Santa Cruz Biotechnology (1 mentions)
Ac 15 antibody, by Merck Group (1 mentions)
Ab54460, by Abcam (1 mentions)
Ab22604, by Abcam (1 mentions)
Ab97046, by Abcam (1 mentions)
Sc 535, by Santa Cruz Biotechnology (1 mentions)
Immobilon fl membrane, by Merck Group (1 mentions)
Sc 8992, by Santa Cruz Biotechnology (1 mentions)
Sc 7273, by Santa Cruz Biotechnology (1 mentions)
Sc 366, by Santa Cruz Biotechnology (1 mentions)
Sc 7975 r, by Santa Cruz Biotechnology (1 mentions)
Trans blot system, by Bio-Rad (1 mentions)
Ac 15, by Merck Group (1 mentions)
X ray film, by Kodak (1 mentions)
2 protocols using ab393364
1
Western Blot Analysis of Antioxidant Proteins
2
Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins concentration was measured using Lowry method (Lowry et al. 1951 ) and 60 µg of proteins were subjected to electrophoresis on 7.5% SDS-PAGE. After transfer to PVDF membrane (Immobilon-FL membrane, Millipore), using a Transblot system (Bio-Rad Laboratories), the unbound sites were blocked (1 h with 2% non-fat dry milk) and the membranes were probed with specific primary rabbit polyclonal antibodies for anti-phospho-p38 Tyr 182 (1:750) (sc-7975-R, Santa Cruz Biotechnology), anti-p38 MAPK (1:500) (sc-535, Santa Cruz Biotechnology), anti-GR (1:500) (sc-8992, Santa Cruz Biotechnology), anti-11BHSD1 (1:1000) (ab393364, Abcam), anti-H6PDH (1:1000) (sc-67394, Santa Cruz Biotechnology), anti-SREBP1c (1:500) (sc-366, Santa Cruz Biotechnology) and mouse monoclonal anti-PPARG (1:1000) (sc-7273, Santa Cruz Biotechnology). Monoclonal mouse anti-B-actin antibody (1:10,000) (AC-15, Sigma-Aldrich) was used as a loading control. Blots were developed with secondary ECL anti-rabbit IgG HRP-linked whole antibody (1:10,000) (Amersham Pharmacia Biotech) or with anti-mouse IgG HRP-linked whole antibody (1:20,000) (ab97046, Abcam). Signal was developed using enhanced chemiluminescence Downloaded from Bioscientifica.com at 09/07/2024 04:13:17AM via free access (ECL) and densitometry of protein bands on X-ray films (Kodak, Rochester, USA) was performed by Image J software (NIH).
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!