Western blots were performed as previously described elsewhere. Membranes were incubated with the following primary antibodies: anti‐MYBBP1A (Proteintech #14524‐AP, Rosemont, IL, USA ), anti‐PGC1α (Abcam #ab54481, Cambridge, UK), anti‐SGLT1 (Abcam #ab14685), anti‐p38 MAPK (Cell Signaling #9212, Danvers, MA, USA), and anti‐phospho‐p38 MAPK (T180/Y182) (Cell Signaling #9215). α‐Tubulin (Sigma #T9026) was used as a loading control. Horseradish peroxidase‐labeled rabbit anti‐mouse (Abcam #ab 97046) and goat anti‐rabbit (Abcam #ab 97051) secondary antibodies were used. The proteins were detected using an ECL detection system (Amersham Biosciences) and Bio‐Rad ChemiDoc XRST (Berkeley, CA, USA).
Chemidoc xrst
Manufactured by Bio-Rad
Sourced in United States
The ChemiDoc XRST is a versatile imaging system designed for capturing and analyzing images of proteins, nucleic acids, and other biomolecules. The device utilizes a high-resolution camera, a range of illumination options, and advanced software to provide accurate, reproducible, and quantitative results.
Automatically generated - may contain errors
Lab products found in correlation
Ecl detection system, by Cytiva (2 mentions)
Ab97046, by Abcam (2 mentions)
Anti mybbp1a, by Proteintech (2 mentions)
α tubulin, by Merck Group (1 mentions)
Anti pgc 1α, by Abcam (1 mentions)
Anti sglt1, by Abcam (1 mentions)
Anti p38 mapk, by Cell Signaling Technology (1 mentions)
Anti phospho p38 mapk t180 y182, by Cell Signaling Technology (1 mentions)
Polyvinylidene fluoride membrane, by Pall Corporation (1 mentions)
Coomassie brilliant blue r 250, by Merck Group (1 mentions)
Ab18184, by Abcam (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Prism ultra protein ladder, by Abcam (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
Anti c myc antibody, by Cell Signaling Technology (1 mentions)
Anti bax antibody, by Cell Signaling Technology (1 mentions)
Anti β actin antibody, by Cell Signaling Technology (1 mentions)
Bicinchoninic acid protein assay kit, by Beyotime (1 mentions)
T9026, by Merck Group (1 mentions)
4 protocols using chemidoc xrst
1
Western Blot Analysis of Cell Signaling
2
SDS-PAGE analysis of purified proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Crude lysates as well as purified proteins were separated in the presence of SDS using 15% polyacrylamide gel and visualized with Coomassie® Brilliant blue R 250 (Merck). Approximately, 30-50 ug/lane purified protein was electrophoresed by SDS-PAGE using Prism Ultra Protein Ladder (ab116028, abcam; 10–245 kDa) and transferred to polyvinylidene fluoride membrane (Pall Corporation, Port Washington, USA). Blots were treated with primary (mouse monoclonal antibody directed against 6X His tag; ab18184, Abcam) and secondary antibodies and incubated with elctrochemiluminescence (ECL) reagent (SuperSignal West Pico Chemiluminescent Substrate, Thermo Scientific) and examined on Bio-Rad-Molecular imager ChemiDoc™ XRSt with ImageLabTM software.
3
Western Blot Analysis of Breast Cancer Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Breast cancer cells or tumor tissues were lysed in the presence of a protease inhibitor cocktail and phosphatase inhibitor mixture (3 µL) and then homogenized. Protein concentration was measured by using a bicinchoninic acid protein assay kit (P0010, Beyotime). Separated proteins were transferred to nitrocellulose membranes and subsequently blocked in 5% nonfat milk solution and incubated with indicated primary antibodies at 4°C overnight (1:2000 dilution; Cell Signaling Technology, Danvers, MA, USA): anti-β-actin antibody, anti-c-Myc antibody, anti-Bax antibody, anti-Bcl-2 antibody, and anti-AKT2 antibody. The membranes then were incubated with the secondary antibody, rabbit horseradish peroxidase-conjugated anti-goat IgG (1:2000 dilution; Beyotime, Hangzhou, China), for 1 h at room temperature. Blotted membranes were detected by using the ChemiDoc XRST and processed by Image Lab Software (both, Bio-Rad, Inc., Hercules, CA, USA). The gray value of each band in the imaging data was analyzed by Quantity One software (Bio-Rad, Inc.).
4
Immunoblotting Protocols for Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blots were performed as previously described elsewhere. Membranes were incubated with the following primary antibodies: anti-MYBBP1A (Proteintech #14524-AP), anti-c-MYB (Millipore #05-175, Billerica, MA, USA), anti-pVHL (Santa Cruz # sc-5575, Santa Cruz, CA, USA), anti-p53 (Santa Cruz #sc-6243), anti-acetyl-p53 (Lys 382) (Cell Signaling #2525, Danvers, MA, USA). a-Tubulin (1:10,000, Sigma #T9026) was used as a loading control. Horseradish peroxidase-labeled rabbit anti-mouse (Abcam # ab 97046) and goat anti-rabbit (Abcam # ab 97051) secondary antibodies were used. The proteins were detected using an ECL detection system (Amersham Biosciences, GE Healthcare, Buckinghamshire, UK) and Bio-Rad ChemiDoc XRST (Hercules, CA, 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!