Whole-cell protein extracts were prepared from samples lysed in RIPA buffer (150 mM NaCl, 5 mM EDTA, 50 mM Tris Base, 1% NP-40, 0.5% Sodium Deoxycholate, 0.1% Sodium Dodecyl Sulfate). Extracts were separated by electrophoresis on NuPAGE 4–12% Bis-Tris gels (Thermo Fisher Scientific) in MOPS buffer (Thermo Fisher Scientific) and blotted onto nitrocellulose membrane (GE Healthcare). Immunoblots were incubated overnight with polyclonal rabbit anti-FMRP antibody (ab27455 Abcam, 1:1000) and for 1 h with monoclonal mouse anti-GAPDH antibody (sc-32233 Santa Cruz, 1:3000) as loading control. Membranes were then incubated with HRP-conjugated donkey anti-mouse IgG (H + L) (IS20404 Immunological Sciences, 1:5000) or donkey anti-rabbit IgG (H + L) (IS20405 Immunological Sciences, 1:5000) secondary antibodies for 45 min. HRP signal was revealed with the Clarity Western ECL Substrates kit (Bio-rad). Images were acquired with the Chemidoc MP (Bio-Rad) and analyzed with the ImageLab software (Bio-Rad).
Clarity western ecl substrates kit
Manufactured by Bio-Rad
The Clarity Western ECL Substrates kit is a chemiluminescent detection system used for the visualization of proteins in Western blot analysis. The kit contains the necessary reagents for the detection of horseradish peroxidase (HRP)-labeled proteins on a membrane.
Automatically generated - may contain errors
Lab products found in correlation
Image lab software, by Bio-Rad (2 mentions)
Nitrocellulose membrane, by GE Healthcare (2 mentions)
Mops buffer, by Thermo Fisher Scientific (2 mentions)
Chemidoc mp, by Bio-Rad (2 mentions)
Nupage 4 12 bis tris gel, by Thermo Fisher Scientific (2 mentions)
Hrp conjugated donkey anti mouse igg h l, by Immunological Sciences (2 mentions)
Donkey anti rabbit igg h l, by Immunological Sciences (2 mentions)
Sc 32233, by Santa Cruz Biotechnology (2 mentions)
2 protocols using clarity western ecl substrates kit
1
Western Blotting Analysis of FMRP
2
Western Blot Analysis of FMRP Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-cell protein extracts were prepared from samples lysed in RIPA buffer (150 mM NaCl, 5 mM EDTA, 50 mM Tris Base, 1% NP-40, 0.5% Sodium Deoxycholate, 0.1% Sodium Dodecyl Sulfate).
Extracts were separated by electrophoresis on NuPAGE 4-12% Bis-Tris gels (Thermo Fisher Scientific) in MOPS buffer (Thermo Fisher Scientific) and blotted onto nitrocellulose membrane (GE Healthcare). Immunoblots were incubated overnight with polyclonal rabbit anti-FMRP antibody (ab27455 Abcam, 1:1000) and for 1 hour with monoclonal mouse anti-GAPDH antibody (sc-32233 Santa Cruz, 1:3000) as loading control. Membranes were then incubated with HRP-conjugated donkey anti-mouse IgG (H+L) (IS20404 Immunological Sciences, 1:5000) or donkey anti-rabbit IgG (H+L) (IS20405 Immunological Sciences, 1:5000) secondary antibodies for 45 minutes. HRP signal was revealed with the Clarity Western ECL Substrates kit (Bio-rad). Images were acquired with the Chemidoc MP (Bio-Rad) and analyzed with the ImageLab software (Bio-Rad).
Extracts were separated by electrophoresis on NuPAGE 4-12% Bis-Tris gels (Thermo Fisher Scientific) in MOPS buffer (Thermo Fisher Scientific) and blotted onto nitrocellulose membrane (GE Healthcare). Immunoblots were incubated overnight with polyclonal rabbit anti-FMRP antibody (ab27455 Abcam, 1:1000) and for 1 hour with monoclonal mouse anti-GAPDH antibody (sc-32233 Santa Cruz, 1:3000) as loading control. Membranes were then incubated with HRP-conjugated donkey anti-mouse IgG (H+L) (IS20404 Immunological Sciences, 1:5000) or donkey anti-rabbit IgG (H+L) (IS20405 Immunological Sciences, 1:5000) secondary antibodies for 45 minutes. HRP signal was revealed with the Clarity Western ECL Substrates kit (Bio-rad). Images were acquired with the Chemidoc MP (Bio-Rad) and analyzed with the ImageLab software (Bio-Rad).
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!