Immunoblotting was performed as described (60 (link)). Briefly, cultured cells were lysed using radioimmunoprecipitation assay buffer supplemented with a protease inhibitor cocktail (Roche). Proteins were resolved with SDS–polyacrylamide gel electrophoresis, and after transfer to a nitrocellulose membrane were blocked in Odyssey blocking buffer (LI-COR). The following primary antibodies were used: human NAF1 (rabbit, ab157106, 1:1000; Abcam), mouse Naf1 (rabbit, Naf1 394, 1:250; Prosci), Myc (mouse, clone 4A6, 1:1000; Millipore), human dyskerin (rabbit, sc-48794, 1:250; Santa Cruz Biotechnology), and green fluorescent protein (mouse, 7.1 and 13.1, 1:1000; Roche) with loading controls actin (mouse, ab8226, 1:2000; Abcam), tubulin (rabbit, ab6046, 1:5000; Abcam), PARP (rabbit, 9542S, 1:1000; Cell Signaling Technology), or GAPDH (mouse, mAbcam9498, 1:1000; Abcam). Secondary antibodies were conjugated to dyes IR680 or IR800 (donkey, 1:10,000; LI-COR), and blots were visualized using an Odyssey scanner (LI-COR), with exception of the mouse Naf1 antibody that was visualized by horseradish peroxidase–linked antibody (rabbit, 7074, 1:10,000; Cell Signaling Technology) and enhanced chemiluminescent substrate (Thermo Scientific). Cell fractionation was performed using NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific).
Ab157106
Manufactured by Abcam
Sourced in United States
Ab157106 is a primary antibody that targets the human EGFR protein. It is a rabbit monoclonal antibody. The antibody is designed for use in applications such as Western blot and Immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions)
Ab8226, by Abcam (1 mentions)
Enhanced chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Ab6046, by Abcam (1 mentions)
Ir800, by LI COR (1 mentions)
Ir680, by LI COR (1 mentions)
Clone 4a6, by Merck Group (1 mentions)
Sc 48794, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase linked antibody, by Cell Signaling Technology (1 mentions)
Odyssey blocking buffer, by LI COR (1 mentions)
Odyssey scanner, by LI COR (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Ab40794, by Abcam (1 mentions)
Ab32417, by Abcam (1 mentions)
Ab81298, by Abcam (1 mentions)
Ab52167, by Abcam (1 mentions)
Ab199380, by Abcam (1 mentions)
Ab32027, by Abcam (1 mentions)
Ab59362, by Abcam (1 mentions)
Ab201172, by Abcam (1 mentions)
2 protocols using ab157106
1
Immunoblotting Procedure for Protein Expression
2
Western Blot Analysis of Protein Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cell and tissue lysates were prepared in 1× sodium dodecyl sulphate (SDS) buffer. Identical quantities of protein were separated by SDS gel electrophoresis and transferred onto nitrocellulose filter membranes. After incubating with antibodies specific for QPCT (ab201172, Abcam, CA, USA) and GAPDH (sc-25778; Santa Cruz, CA, USA), the blots were incubated with IRDye 800-conjugated goat anti-rabbit IgG, and bands were detected using an Odyssey infrared scanner (Li-Cor). GAPDH was used as the loading control. The other antibodies used for western blot were against IRS1 (ab52167, Abcam, CA, USA), NF-κB (p65) (8242, Cell Signalling Technology), HRAS (ab32417, Abcam, CA, USA), CBL (ab32027, Abcam, CA, USA), GAB1 (ab59362, Abcam, CA, USA), NAF1 (ab157106, Abcam, CA, USA), MAPK8 (ab199380, Abcam, CA, USA), MAPK10 (ab126591, Abcam, CA, USA), FAK (PTK2) (ab40794, Abcam, CA, USA), p-FAK (ab81298, Abcam, CA, USA), ERK1/2 (4695, Cell Signaling Technology), p-ERK1/2 (4370, Cell Signaling Technology), AKT (4691, Cell Signaling Technology), p-AKT (4060, Cell Signaling Technology), Stat3 (9139, Cell Signaling Technology), p-Stat3 (9145, Cell Signaling Technology), and ubiquitin (3936, Cell Signaling Technology).
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!