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Product name
NADH Dehydrogenase (Complex I) Profiling ELISA Kit
See all Complex I products (7) ...
Specificity
Species– Human, Bovine. Rat and Mouse samples are not appropriate, other species are untested.
Precision
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| Overall | 4 | 4.2% |
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| Overall | 3 | 6.2% |
Tests
1 x 96 test
Sample type
Tissue Extracts, Cell Lysate
Assay type
Sandwich
Sensitivity
20 µg/ml
Recovery
| Sample type | Average % | Range % |
|---|---|---|
| Tissue Extracts | 3 - 200 | |
| Cell Lysate | 20 - 1000 |
Cross reactivity
Reacts with
Cow, Human
Product overview
NADH dehydrogenase (Complex I) is the first enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. NADH dehydrogenase is a large protein complex of 950,000 MW made up of 45-46 different subunits. Seven of the subunits of the complex are encoded on mitochondrial DNA (mtDNA), the remaining subunits are nuclear encoded, made in the cytosol and translocated into the organelle for assembly at the inner membrane. The enzyme complex catalyses electron entry from NADH via a flavin (FMN) and several non-heme iron centers.
ab124535 NADH dehydrogenase (Complex I) Human Profiling Kit is an in vitro enzyme-linked immunosorbent assay (ELISA) for the comparison of NADH dehydrogenase levels or profile in cell and tissue lysates. The assay employs an NADH dehydrogenase specific antibody coated onto microplate well plate strips.
Samples are pipetted into the wells and NADH dehydrogenase present in the sample is bound to the wells by the immobilized antibody. The wells are washed and a biotin labelled anti-NADH dehydrogenase detector antibody is added. After washing away unbound detector antibody, biotin affinity HRP-conjugated streptavidin is pipetted into the wells. The wells are again washed, an HRP substrate solution (TMB) is added to the wells and color develops in proportion to the amount of NADH dehydrogenase (Complex I) bound. The developing blue color is measured at 600 nm.
Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.
Notes
Store all components at 4°C. This kit is stable for 6 months from receipt. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.
Tested applications
Sandwich ELISAmore details
Properties
Storage instructions
Please see Notes section
| Components | Identifier | 1 x 96 tests |
|---|---|---|
| 10X HRP Label | 8206020 | 1 x 1ml |
| 10X Blocking Buffer | 8209802 | 1 x 6ml |
| 10X NADH Dehydrogenase Detector Antibody | 8209559 | 1 x 1ml |
| 20X Buffer | 8203013 | 1 x 20ml |
| Extraction Buffer | 8201093 | 1 x 15ml |
| NADH Dehydrogenase Microplate | 8209560 | 1 unit |
| TMB Development Solution | 8209803 | 1 x 12ml |
Alternative names
- MS1035
Applications
Show applications key
Our Abpromise guarantee covers the use of ab124535 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
sELISA
NADH Dehydrogenase (Complex I) Profiling ELISA Kit images:
Sandwich ELISA - NADH Dehydrogenase (Complex I) Profiling ELISA Kit (ab124535)
Example control sample curve of serially titrated HepG2 cell extracts in the working range of the assay.
Sandwich ELISA - NADH Dehydrogenase (Complex I) Profiling ELISA Kit (ab124535)
Human HepG2 cells were cultured in NARTI Zalcitabine (ddC) for 6 days to ensure a significant effect on mitochondrial DNA replication. The drug reduced mitochondrial DNA levels and hence mitochondrial protein expression. As a consequence the assembly of Complexes I, III and IV were severely affected. Note that loss of the two small mitochondrial DNA encoded subunits of Complex V (ATP synthase) does not affect overall assembly. Interestingly an increase in Complex II was induced as a consequence of I, III, IV loss possibly to up regulate mitochondrial citric acid cycle function.
Sandwich ELISA - NADH Dehydrogenase (Complex I) Profiling ELISA Kit (ab124535)
Human HepG2 cells were cultured in chloramphenical for 6 days to ensure a significant effect on mitochondrial protein translation. The antibiotic inhibited mitochondrial protein translation and assembly of Complexes I and IV but had no significant effect on Complex III.
Protocols
References for NADH Dehydrogenase (Complex I) Profiling ELISA Kit (ab124535)
ab124535 has not yet been referenced specifically in any publications.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"