Usingclarity western ecl substrate

Manufactured by Bio-Rad

UsingClarity Western ECL Substrate is a chemiluminescent detection reagent designed for the quantification of proteins on Western blots. It produces a luminescent signal that can be detected and quantified using a compatible imaging system.

Automatically generated - may contain errors

Lab products found in correlation

Ab174318, by Abcam (1 mentions) Ab58345, by Abcam (1 mentions) Thechemidoc mp, by Bio-Rad (1 mentions) Trans blot turbo, by Bio-Rad (1 mentions) Bis tris gel, by Thermo Fisher Scientific (1 mentions) Anti mouse hrp conjugate, by Merck Group (1 mentions) Lds sample buffer, by Thermo Fisher Scientific (1 mentions) Mouse monoclonal anti flag m2 antibody, by Merck Group (1 mentions) Image lab software, by Bio-Rad (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions)

2 protocols using usingclarity western ecl substrate

Western Blot Analysis of Protein Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein samples were loaded onto an SDS-PAGE gel and transferred to
a PVDF membrane using Trans-Blot Turbo (Bio-Rad, 170-4273) following the
manufacture’s protocol. Western blotting signals were developed using
Clarity Western ECL Substrate (Bio-Rad, 170-5061) and imaged with the
ChemiDoc MP (Bio-Rad, 17001402). Primary antibody dilutions were: 1:1000 for
Met-14-3-3γ (Novus, NB100-407), 1:1000 for
14-3-3γ (Cell Signaling, D15B7), 1:250 for HA
(Thermo Fisher, SG77), 1:1000 for Rps5/uS7 (Abcam, ab58345), 1:1000 for Zuo1
(Dr. Elizabeth Craig) (Lee et al.,
2016), 0.4 μg/ml for bacteria Rps13/uS13 (Developmental
Studies Hybridoma Bank, 193E11E5B11), 1:500 for MetAP1 (R&D, MAB3537),
1:1000 for MetAP2 (Proteintech, 17040-1-AP), 1/1000 for Rpl10a/uL1 (Abcam,
ab174318).

Fujii K., Susanto T.T., Saurabh S, & Barna M. (2018). Decoding the Function of Expansion Segments in Ribosomes. Molecular cell, 72(6), 1013-1020.e6.

+ Open protocol

+ Expand

Western Blot Quantification of Protein Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein samples were
reduced and denatured by boiling in LDS sample buffer (Life Technologies)
containing 1 mM 2-mercaptoethanol for 10 min prior to resolution on
4–12% Bis-Tris gels (Life Technologies). Under these conditions,
no additional intein cleavage above that of the 20-h MESNA reaction
is observed. Proteins were subsequently transferred to a nitrocellulose
membrane for Western blot analysis. Detection of FLAG tagged proteins
was performed by probing the membranes with anti-FLAG M2 mouse monoclonal
antibody (Sigma-Aldrich, diluted 1:3000) followed by anti-mouse HRP
conjugate (Sigma-Aldrich, diluted 1:2000). To detect biotinylated
proteins, membranes were probed with anti-biotin mouse monoclonal
antibody Ab-2 clone BTN.4 (Lab Vision Corporation, diluted 1:500)
followed by anti-mouse HRP conjugate. Membranes were developed using
Clarity Western ECL Substrate (Bio-Rad) and imaged with the ChemiDoc
XRS+ system (Bio-Rad). Unsaturated band intensities were measured
with the Image Lab Software (Bio-Rad) to quantify the relative protein
amounts.

Marshall C.J., Grosskopf V.A., Moehling T.J., Tillotson B.J., Wiepz G.J., Abbott N.L., Raines R.T, & Shusta E.V. (2014). An Evolved Mxe GyrA Intein for Enhanced Production of Fusion Proteins. ACS Chemical Biology, 10(2), 527-538.

+ 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?

Revolutionizing how scientists
search and build protocols!