Immunoblot analysis were performed as previously described [19 (link)]. Anti-Casp1 p10, anti-IL-18, anti-desmin (all three from Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-IL-1β (Abcam, Cambridge, MA, USA), and anti-alpha smooth muscle actin (α-SMA; GeneTex, Irvine, CA, USA) antibodies were used in combination with appropriate peroxidase-conjugated secondary antibodies. Protein load was verified with an α-tubulin antibody (dilution 1:10,000) (Hybridomabank, University of Iowa) (kindly provided by M. Kaulich). Bands were visualized with the enhanced chemiluminescence reagent and digitized using a CCD camera (ChemiDoc®, Bio-Rad, Hercules, CA, USA). Expression intensity was quantified by ImageLab (Bio-Rad).
Protocol detail
Immunoblot Analysis of Cellular Signaling
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Alpha actin
Antibodies
Antibody
Chemiluminescence
Desmin
Dilution
Il 18
Il 1β
Immunoblot analysis
Peroxidase
Protein
Smooth muscle
α tubulin
Corresponding Organization :
Other organizations : University of California, San Diego, Essen University Hospital, University of Buenos Aires, Ludwig Cancer Research
Top 5 similar protocols
Variable analysis
independent variables
- Anti-Casp1 p10 antibody
- Anti-IL-18 antibody
- Anti-desmin antibody
- Anti-IL-1β antibody
- Anti-alpha smooth muscle actin (α-SMA) antibody
- Protein expression levels
- Protein load verified with an α-tubulin antibody (dilution 1:10,000)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!