Total proteins were prepared using lysis buffer (Cell Signaling Technologies, Danvers, MA) and resolved by SDS–PAGE electrophoresis. Proteins were transferred to nitrocellulose membrane using the iBlot system (Life Technologies, Calrsbad, CA) and blocked with Odyssey Blocking Buffer (Li-Cor Biosciences, Lincoln, NE). The membranes were probed with the following primary antibodies, using the indicated dilution: rabbit anti-GK (1/500), goat anti-PFK (1/1000), anti-PEPCK (1/1000), anti FBPase (1/500), rabbit anti-pAMPK (1/500), mouse anti-AMPK (1/1000), rabbit anti-FOXO1 (1/500), mouse anti-tubulin (1/5000), rabbit anti-PDK4 (1/500), mouse anti-PDH (1/1000), rabbit anti-pPDH (1/500), and goat anti-GKRP (1/2000). This was followed by treatment with infrared IR-dye 700CW and/or 800CW-labeled secondary antibodies (Li-Cor Biosciences, Lincoln, NE). The blots were then scanned and protein levels quantified using Li-Cor Odyssey infrared imaging instrument and Odyssey 2.1 software.
800cw labeled secondary antibodies
Manufactured by LI COR
The 800CW-labeled secondary antibodies are fluorescently-labeled reagents designed for use in immunoassays and other detection methods. They are manufactured to high quality standards and provide consistent performance.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey infrared imaging instrument, by LI COR (3 mentions)
Odyssey blocking buffer, by LI COR (3 mentions)
Lysis buffer, by Cell Signaling Technology (2 mentions)
Iblot system, by Thermo Fisher Scientific (2 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
Odyssey 2, by LI COR (1 mentions)
3 protocols using 800cw labeled secondary antibodies
1
Quantitative Protein Expression Analysis
2
Protein Quantification and Immunoblot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein was isolated using cell lysis buffer (Cell Signaling Technologies, Danvers, MA), as previously described [8 (link)]. Briefly, 40μg of protein per lane was resolved by SDS—PAGE, followed by transfer to nitrocellulose membrane and blocked with Odyssey Blocking Buffer (Li-Cor Biosciences, Lincoln, NE). The membranes were probed with the following primary antibodies, using the indicated dilutions: rabbit anti-HK (1/500), goat anti-PFK (1/1000), rabbit anti-PK (1/1000), rabbit anti-pAMPK (1/500), rabbit anti-pAkt (1/1000), rabbit anti-FOXO1 (1/500), mouse anti-tubulin (1/5000) and rabbit anti-PDK4 (1/500). This was followed by treatment with infrared IR-dye 700CW and/or 800CW-labeled secondary antibodies (Li-Cor Biosciences, Lincoln, NE). Blots were scanned and quantified using Li-Cor Odyssey infrared imaging instrument and Odyssey 2.1 software.
3
Quantitative Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were prepared using lysis buffer (Cell Signaling Technologies, Danvers, MA) and resolved by SDS–PAGE electrophoresis. Proteins were transferred to nitrocellulose membrane using the iBlot system (Life Technologies, Calrsbad, CA) and blocked with Odyssey Blocking Buffer (Li-Cor Biosciences, Lincoln, NE). The membranes were probed with the following primary antibodies, using the indicated dilution: rabbit anti-GK (1/500), goat anti-PFK (1/1000), anti-PEPCK (1/1000), anti FBPase (1/500), rabbit anti-pAMPK (1/500), mouse anti-AMPK (1/1000), rabbit anti-FOXO1 (1/500), mouse anti-tubulin (1/5000), rabbit anti-PDK4 (1/500), mouse anti-PDH (1/1000), rabbit anti-pPDH (1/500), and goat anti-GKRP (1/2000). This was followed by treatment with infrared IR-dye 700CW and/or 800CW-labeled secondary antibodies (Li-Cor Biosciences, Lincoln, NE). The blots were then scanned and protein levels quantified using Li-Cor Odyssey infrared imaging instrument and Odyssey 2.1 software.
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