Crude muscle protein extracts were obtained by homogenization of weighted frozen tissue with a buffer solution (62.5 mM Tris, pH 6.8, 2% [w/v] SDS, 5% [v/v] glycerol, 0.10 mM SERVA blue, 0.25 mM EDTA, 30 μM phenol red and 0.9% β-mercaptoethanol) in liquid nitrogen. The samples were incubated for 5 min at 100 • C and centrifuged for 5 min at 20,000 × g. Samples were diluted at 500 μg/well and loaded into 12 or 15% polyacrylamide gels. Nitrocellulose electro blotted membranes (Amersham Biosciences, Uppsala, Sweden) were probed with 1.5 μg/mL chicken polyclonal anti-Gal-3; all commercial antibodies were used according to manufacturers' instructions and were then incubated with 40 ng/mL of a specific HRP conjugate secondary antibody. Bounded antibodies were revealed by enhanced chemiluminescence using the Luminata Forte HRP Substrate (Millipore, Billerica, MA). We used desmin as a constitutive muscle tissue marker (Costa et al. 2004; Cerri et al. 2008) .
Nitrocellulose electro blotted membranes
Manufactured by Cytiva
Sourced in Sweden
Nitrocellulose electro blotted membranes are a type of laboratory equipment used for protein transfer and analysis. They provide a support medium for the immobilization of proteins during the Western blotting process. The membranes are made of nitrocellulose, a material that binds proteins efficiently, enabling their detection and analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nitrocellulose electro blotted membranes
1
Muscle Protein Extraction and Western Blot
2
Muscle Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Crude muscle protein extracts were obtained by homogenization of weighted frozen tissue with a buffer solution (62.5 mM Tris, pH 6.8, 2% (w/v) SDS, 5% (v/v) glycerol, 0.10 mM SERVA blue, 0.25 mM EDTA, 30 μM phenol red, and 0.9% β-mercaptoethanol) in liquid nitrogen. The samples were incubated for 5 min at 100°C and centrifuged for 5 min at 20,000 x g. Samples were diluted at 500 μg/well and loaded into 12 or 15% polyacrylamide gels. Nitrocellulose electro blotted membranes (Amersham Biosciences, Uppsala, Sweden) were probed with 1.5 μg/mL chicken polyclonal anti-Gal-3; all commercial antibodies were used in according to manufacturers' instructions; and then incubated with 40 ng/mL of a specific HRP conjugate secondary antibody.
Bounded antibodies were revealed by enhanced chemiluminescence using the Luminata Forte HRP Substrate (Millipore, Billerica, MA).
Bounded antibodies were revealed by enhanced chemiluminescence using the Luminata Forte HRP Substrate (Millipore, Billerica, MA).
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!