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For identifying inhibitors and activators of mitochondrial biogenesis in adherent cultured cells. Each kit contains sufficient reagents to analyze two 96-well plates of fixed human, rat, mouse, or bovine cells. This kit utilizes IRDyes® for detection, and so requires a LI-COR® Odyssey® or Aerius® imaging system. An alternate colorimetric version of this kit is available for use with standard plate readers - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217/MS643).
In-Cell ELISA Kits use quantitative immunocytochemistry to measure protein levels or post-translational modifications in cultured cells. Cells are fixed in a 96-well plate and targets of interest are detected with highly-specific, well-characterized monoclonal antibodies, and levels are quantified with IRDye®-labeled Secondary Antibodies. IR imaging and quantitation is performed using a LI-COR® Odyssey® or Aerius® system.
ab110215 (MS642) is designed to measure drug-induced effects on mitochondrial biogenesis early in the safety screening process. The MitoBiogenesis™ In-Cell ELISA Kit is a true duplexing 96/384-well assay that ratios both an mtDNA- and an nDNA-encoded protein in cultured or primary cells, and which requires very little sample prep and few overall steps.
Cells (human, rat or mouse) are seeded in 96- or 384-well microplates, and after exposure to experimental compounds for several cell doublings, the levels of two mitochondrial proteins are measured simultaneously in each well. The two proteins are each subunits of a different oxidative phosphorylation enzyme complex, one protein being subunit I of Complex IV (COX-I), which is mtDNA-encoded, and the other being the 70 kDa subunit of Complex II (SDH-A), which is nDNA-encoded. Complex IV includes several proteins which are encoded in the mitochondrion, while the proteins of Complex II are entirely encoded in the nucleus. Optionally, total protein levels can also be measured.
LI-COR®, Odyssey®, Aerius®, IRDye™ and In-Cell Western™ are registered trademarks or trademarks of LI-COR Biosciences Inc
Plates are available in our ICE (In-Cell ELISA) Support Pack (ab111542) which can be bought seperately
|Components||2 x 96 tests|
|1000X IRDye-labeled Secondary Antibodies||1 x 24µl|
|100X Triton X-100||1 x 0.5ml|
|10X Blocking Buffer||1 x 15ml|
|10X Phosphate Buffered Saline||1 x 100ml|
|200X Primary Antibodies||1 x 0.1ml|
|400X Tween-20||1 x 2ml|
|Janus Green Stain||1 x 11ml|
|Plate Seals||2 units|
Our Abpromise guarantee covers the use of ab110216 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|In-Cell ELISA||Use at an assay dependent dilution.|