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MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) is an in-cell ELISA used to identify inhibitors and activators of mitochondrial biogenesis in adherent cultured cells.

- Colorimetric readout - works on any standard plate reader
- Works in 96-well or 384-well plates for higher throughput
- Designed to measure drug-induced effects on mitochondrial biogenesis early in the safety screening process.


Images

Western blot - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (AB110217), expandable thumbnail
  • Immunocytochemistry - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (AB110217), expandable thumbnail
  • In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (AB110217), expandable thumbnail
  • In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (AB110217), expandable thumbnail
  • In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (AB110217), expandable thumbnail

Publications

Key facts

Detection method
Colorimetric
Sample types
Adherent cells
Assay type
Cell-based (quantitative)
Reactive species
Mouse, Rat, Cow, Human

Reactivity data

Application
ELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Flavoprotein (FP) subunit of succinate dehydrogenase (SDH) that is involved in complex II of the mitochondrial electron transport chain and is responsible for transferring electrons from succinate to ubiquinone (coenzyme Q) (PubMed:10746566, PubMed:24781757). SDH also oxidizes malate to the non-canonical enol form of oxaloacetate, enol-oxaloacetate (By similarity). Enol-oxaloacetate, which is a potent inhibitor of the succinate dehydrogenase activity, is further isomerized into keto-oxaloacetate (By similarity). Can act as a tumor suppressor (PubMed:20484225).

Additional Targets

MT-CO1

Alternative names

SDH2, SDHF, SDHA, Flavoprotein subunit of complex II, Malate dehydrogenase [quinone] flavoprotein subunit, Fp

What's included?

2 x 96 Tests
Components
100X Triton X-100
1 x 1.5 mL
10X Blocking Buffer
1 x 15 mL
10X Phosphate Buffered Saline
1 x 100 mL
1X AP Development Solution
1 x 24 mL
1X HRP Development Solution
1 x 24 mL
1X Janus Green Stain
1 x 17 mL
200X Primary Antibodies
1 x 0.1 mL
2500X AP-labeled Secondary Antibody
1 x 12 µL
2500X HRP-labeled Secondary Antibody
1 x 12 µL
400X Tween-20
1 x 2 mL
AP Development Reagent
1 x 139 mg
Plate Seals
2 x 1 Unit

Recommended products

MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) is an in-cell ELISA used to identify inhibitors and activators of mitochondrial biogenesis in adherent cultured cells.

- Colorimetric readout - works on any standard plate reader
- Works in 96-well or 384-well plates for higher throughput
- Designed to measure drug-induced effects on mitochondrial biogenesis early in the safety screening process.

Key facts

Detection method
Colorimetric
Sample types
Adherent cells
Assay type
Cell-based (quantitative)
Reactive species
Mouse, Rat, Cow, Human
Assay Platform
Microplate

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

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 colorimetric detection for use with standard plate readers. An alternate IR version of this kit is available which utilizes LI-COR® near-infrared IRDyes® for detection - MitoBiogenesis™ In-Cell ELISA Kit (IR) (MitoBiogenesis™ In-Cell ELISA Kit (IR) ab110216/MS642).

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 enzyme-labeled secondary antibodies.

ab110217 MitoBiogenesis™ In-Cell ELISA Kit (ab110217/MS643) is designed to measure drug-induced effects on mitochondrial biogenesis early in the safety screening process. ab110217 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 70kDa 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® and IRDye® are registered trademarks of LI-COR Biosciences Inc

Plates are available in our ICE (In-Cell ELISA) Support Pack (In-Cell ELISA (ICE) Support Pack ab111542) which can be bought separately.

**Related products** Review the , or the full to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Western blot - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217), expandable thumbnail

    Western blot - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217)

    Antibody specificity demonstrated by Western Blot. A Western blot of total cell protein (10 µg) from human or rat cultured cells was probed with the primary and secondary antibodies and scanned with a LI-COR® Odyssey® imager. The two mitochondrial proteins targeted by the two primary mAbs were labeled and visualized specifically despite the presence of thousands of other proteins. Furthermore, reduction of mtDNA levels in human Rho0 (mtDNA-depleted) cells, or inhibition of mitochondrial protein translation by chloramphenicol in rat cells both result in specific reduction of COX-I protein while nuclear DNA-encoded SDH-A is unaffected.

  • Immunocytochemistry - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217), expandable thumbnail

    Immunocytochemistry - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217)

    Antibody specificity demonstrated by immunocytochemistry. Two-color immunocytochemical labeling of cultured cells with the two MS643 primary monoclonal antibodies specific for COX-I and SDH-A. The two antibodies exhibit striking and specific co-localization in the mitochondria, consistent with the known mitochondrial expression of both proteins.

  • In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217), expandable thumbnail

    In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217)

    Quantitative measurement of the COX-I/SDH-A protein expression ratio. At all cell concentrations, a consistent ratio of mtDNA-encoded protein expression (COX-I) to nuclear DNA-encoded mitochondrial protein expression (SDH-A) is observed in untreated cells. Therefore, normalizing COX-I levels to SDH-A levels simplifies data analysis and eliminates the need to perform all tests at the same cell concentration.

  • In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217), expandable thumbnail

    In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217)

    Inhibition of mitochondrial biogenesis by chloramphenicol The IC50 of a drug's effect on mitochondrial protein translation can be determined quickly using the MitoBiogenesis™ In-Cell ELISA Kit. In this example, cells were seeded at 6000 cells/well, allowed to grow for 3 cell doublings in a drug dilution series and then the relative amounts of COX-I, and SDH-A were measured in each well. Chloramphenicol inhibits mtDNA-encoded COX-I protein synthesis relative to nuclear DNA-encoded SDH-A protein synthesis by 50% at 8.1 µM.

  • In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217), expandable thumbnail

    In-Cell ELISA - MitoBiogenesis™ In-Cell ELISA Kit (Colorimetric) (ab110217)

    Cells are grown to ~80% confluency in a 96- or 384-well plate, a drug/other treatment is applied to stimulate a cellular response. The cells are then fixed and permeabilized, effectively "freezing" them. Primary antibodies are then added which bind to their intended targets within the mitochondria or other subcellular compartment. After incubation, the unbound primary antibodies are washed away and secondary antibodies are added. These secondaries are conjugated to either IRDyes® or to an enzyme label (HRP or AP) for the colorimetric versions of the assays. Unbound secondaries are washed away, reaction buffer is added for the colorimetric assays, and the signal is read on a suitable instrument for the kit type.» In-cell ELISA diagram in PDF format

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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