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Read our guarantee »Overview
Product name
Fumarase Specific Activity Assay Kit
Assay type
Enzyme activity
Cross reactivity
Reacts with
Rat, Cow, Human
Product overview
ab110043 (MS780) is a novel assay that uses a high affinity monoclonal capture antibody to selectively isolate fumarase (FH), which removes all other enzymes of the Krebs cycle and NAD+/NADH utilizing enzymes. After isolation and subsequent measurement of the enzyme's functional activity, the quantity of isolated FH is measured in the same well by adding a second monoclonal detector antibody, which is quantified using a colorimetric label (HRP). Both reactions take place in time-dependent manners proportional to the amount of enzyme captured in each well. By combining activity and quantity measurements, the enzyme's relative specific activity can be determined. Specific activity is useful for measuring up or down regulation of activity by site-specific modification or damage, and in response to specific inhibitors.
Notes
All components are shipped cold. Upon receipt store 200X
Fumarate, 200X bNAD+, 100X Coupler and 100X Reagent Dye at
-20°C or preferably at -80°C for longer stability. Store all other
components at 4°C.
Fumarate, 200X bNAD+, 100X Coupler and 100X Reagent Dye at
-20°C or preferably at -80°C for longer stability. Store all other
components at 4°C.
Tested applications
Sandwich ELISAmore details
Properties
Storage instructions
Please see Notes section
| Components | 96 tests |
|---|---|
| 100X Coupler | 1 x 250µl |
| 100X Detector Antibody | 1 x 130µl |
| 100X HRP Label | 1 x 130µl |
| 100X Reagent Dye | 1 x 250µl |
| 10X Blocking Solution | 1 x 10ml |
| 1X Base Buffer | 1 x 25ml |
| 1X Development Solution | 1 x 12ml |
| 2000X Malate dehydrogenase | 1 x 20µl |
| 200X betaNAD+ | 1 x 250µl |
| 200X Fumarate (1M) | 1 x 250µl |
| 20X Wash Buffer | 1 x 25ml |
| 96-well microplate (12 strips) | 1 unit |
| Extraction Buffer | 1 x 15ml |
Research areas
Metabolism >> Pathways and Processes >> Metabolic signaling pathways >> Energy transfer pathways >> Energy Metabolism
Metabolism >> Pathways and Processes >> Metabolic signaling pathways >> Carbohydrate metabolism
Cancer >> Cancer Metabolism >> Metabolic signaling pathway >> Metabolism of carbohydrates
Kits/ Lysates/ Other >> Kits >> Cell Metabolism Kits >> Other Metabolism Assay
Signal Transduction >> Metabolism >> Energy Metabolism
Applications
Show applications key
Our Abpromise guarantee covers the use of ab110043 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
sELISA
Fumarase Specific Activity Assay Kit images:
Sandwich ELISA - Fumarase Specific Activity Assay Kit (ab110043)
Abcams' protein quantity microplate assays use the well-established sandwich ELISA format, whereby capture and detector antibodies are used to immobilize and then quantify a target protein or enzyme. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state. Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, a second monoclonal antibody, against a different epitope on the target, is added to the well. This detector antibody is either directly labeled with biotin, or a biotin-labeled goat anti-mouse secondary is added. Substrate plus HRP or AP conjugated to streptavidin provide a colorimetric signal that is readable by any plate readers capable of standard ELISA absorbance measurements.
- Fumarase Specific Activity Assay Kit (ab110043)
Figure 1. FH activity and quantity data were collected in 2 control cell lines and a patient cell line (P1). The patient P1 has extra genetic material including a copy of the FH gene. Analysis of this cell line alongside the control cell lines shows that P1 has a greater amount and activity of FH at all sample dilutions. However the specific activity (ratio of activity to quantity) of FH in the patient cell line is very similar to the control cell lines.
Protocols
References for Fumarase Specific Activity Assay Kit (ab110043)
ab110043 has not yet been referenced specifically in any publications.
Publishing research using ab110043? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"