Log In Sign up
The largest database of trusted experimental protocols

Reliablot kit

Manufactured by Fortis Life Sciences
Visit Supplier

The ReliaBLOT kit is a laboratory equipment product designed for Western blot analysis. It provides a reliable and efficient method for the detection and quantification of specific proteins within a sample.

Automatically generated - may contain errors

Lab products found in correlation

Criterion gradient gels, by Bio-Rad (2 mentions) Complete protease inhibitor cocktail, by Merck Group (2 mentions) Odyssey infrared imaging system, by LI COR (2 mentions) Trans blot sd cell, by Bio-Rad (2 mentions) Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions) Nupage tris acetate gel, by Thermo Fisher Scientific (1 mentions) P smad2, by Cell Signaling Technology (1 mentions) Ecl detection system, by GE Healthcare (1 mentions)

Spelling variants of this product

ReliaBLOT (2 citations)

3 protocols using reliablot kit

1

Protein Extraction and Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in 1x RIPA buffer (1% NP-40, 0.1% SDS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% sodium deoxycholate, 1 mM EDTA) for whole cell extracts, or 1x EBC buffer (120 mM NaCl, 0.5% v/v NP-40, 50 mM Tris-HCl pH 8.0) for immunoprecipitations, 0.5 μM DTT (0.01% 2-Mercaptoethanol for EBC buffer), 25 mM NaF, 1 mM sodium orthovanadate, 1 mM PMSF, and cOmplete protease inhibitor cocktail (Sigma). PVDF membranes were blocked in 5% milk/0.1% Tween 20 TBS for 1 hr at room temperature. Primary antibodies were incubated overnight at 4°C. After incubation with either the secondary IRDye Alexa Fluor 680 goat antimouse antibody or 800 goat anti-rabbit antibodies (Odyssey), the membranes were visualized with the Odyssey Infrared Imaging System (Li-Cor). Immunoprecipitations: whole cell lysates (10% inputs) and immunoprecipitates were resolved on 4%–15% gradient Criterion gels (BioRad) and transferred to nitrocellulose membranes using semi-dry Transblot SD cell (BioRad). ReliaBLOT kit (Bethyl Labs; WB120) was used for IP/western blotting according to manufacturer’s protocol and membranes were probed with indicated antibodies.
Bainor A.J., Saini S., Calderon A., Casado-Polanco R., Giner-Ramirez B., Moncada C., Cantor D.J., Ernlund A., Litovchick L, & David G. (2018). The HDAC-Associated Sin3B Protein Represses DREAM Complex Targets and Cooperates with APC/C to Promote Quiescence. Cell reports, 25(10), 2797-2807.e8.
+ Open protocol
+ Expand
2

Immunoblot Analysis of Protein Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissue lysates were resolved in 4-12% or 12% NuPAGE Bis-Tris gels and immunoprecipitates in 7% NuPAGE Tris-Acetate gels (Invitrogen). They were all blotted onto nitrocellulose membrane (Invitrogen) and immunostained. Immunoprecipitates were assayed using ReliaBLOT kit (WB120, Bethyl Laboratories). Antibody dilutions for lung lysates were as follows: FBXO11, 1:2000; p53, 1:500; pSMAD2 (3101, Cell Signaling) 1:500; p21, 1:1000; PAI-1, 1:1000; MMP-2, 1:500; BCL-6, 1:1000; SET8, 1:500; NEDD8, 1:150; and CDT2, 1:250. Antibody dilutions for immunoprecipitates were: FBXO11, 1:400; p53, 1:25; CUL1, 1:100; and CUL4, 1:100. ECL detection system (GE Healthcare) was used.
Tateossian H., Morse S., Simon M.M., Dean C.H, & Brown S.D. (2015). Interactions between the otitis media gene, Fbxo11, and p53 in the mouse embryonic lung. Disease Models & Mechanisms, 8(12), 1531-1542.
+ Open protocol
+ Expand
3

Protein Extraction and Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in 1x RIPA buffer (1% NP-40, 0.1% SDS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% sodium deoxycholate, 1 mM EDTA) for whole cell extracts, or 1x EBC buffer (120 mM NaCl, 0.5% v/v NP-40, 50 mM Tris-HCl pH 8.0) for immunoprecipitations, 0.5 μM DTT (0.01% 2-Mercaptoethanol for EBC buffer), 25 mM NaF, 1 mM sodium orthovanadate, 1 mM PMSF, and cOmplete protease inhibitor cocktail (Sigma). PVDF membranes were blocked in 5% milk/0.1% Tween 20 TBS for 1 hr at room temperature. Primary antibodies were incubated overnight at 4°C. After incubation with either the secondary IRDye Alexa Fluor 680 goat antimouse antibody or 800 goat anti-rabbit antibodies (Odyssey), the membranes were visualized with the Odyssey Infrared Imaging System (Li-Cor). Immunoprecipitations: whole cell lysates (10% inputs) and immunoprecipitates were resolved on 4%–15% gradient Criterion gels (BioRad) and transferred to nitrocellulose membranes using semi-dry Transblot SD cell (BioRad). ReliaBLOT kit (Bethyl Labs; WB120) was used for IP/western blotting according to manufacturer’s protocol and membranes were probed with indicated antibodies.
Bainor A.J., Saini S., Calderon A., Casado-Polanco R., Giner-Ramirez B., Moncada C., Cantor D.J., Ernlund A., Litovchick L, & David G. (2018). The HDAC-Associated Sin3B Protein Represses DREAM Complex Targets and Cooperates with APC/C to Promote Quiescence. Cell reports, 25(10), 2797-2807.e8.
+ 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!