MitoSciences: MSFX-10 FlexPlex™ Module: Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD)

Enzymes > Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) > FlexPlex™ Module: Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD)

FlexPlex™ Module: Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD)

Catalog No. MSFX-10

$125.00 - 1 module

The Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) FlexPlex™ Module is used to determine the quantity of SCHAD (short-chain 3-hydroxyacyl-CoA dehydrogenase) enzyme in a sample. SCHAD is immunocaptured within the wells. The quantity of protein is measured by adding a second SCHAD specific antibody that is then labeled with horseradish peroxidase. This peroxidase changes the substrate from colorless to blue. The rate of color development is proportional to the amount of protein captured in the well and can be monitored at 600 nm. Alternatively, the assay can be terminated, at a user defined time, by the addition of 1N HCl (not supplied) and the assay performed as an end point measurement at 450 nm.

Click here to learn more about short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD).

Specifications

Data

Documentation

Citations

Product Specifications
Applications:	analysis of SCHAD quantity
Species Reactivity:	human
Range (min. 2x background):	0.6 - 20 µg / well (Example: HepG2)
Av. Intra-assay %CV (n=6):	8.9%
Storage Conditions:	Store at 4°C.
Country of Origin:	USA

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Specifications

Data

Documentation

Citations

Figure 1

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Figure 1. HepG2 titration series. Increasing amounts of HepG2 cells were analyzed using MSFX-10 to demonstrate the potential working range of the assay.

Figure 2

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Figure 2. HL60 titration series. Increasing amounts of HL60 were analyzed using MSFX-10 to demonstrate the potential working range of the assay.

Figure 3

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Figure 3. Human heart titration series. Increasing amounts of human heart homogenate were analyzed using MSFX-10 to demonstrate the potential working range of the assay.

Figure 4

	Fibroblasts extract loaded per well	Signal / background (control)	Reproducibility (%CV 6 wells)
MSFX-9 [MCAD]	40 µg	14	4.4
MSFX-10 [SCHAD]	10 µg	224	2.3
MSFX-11 [TFP]	1.25 µg	23	3.0
MSFX-12 [SOD2]	5 µg	4.8	12.2
MSFX-15 [PDH]	5 µg	4.3	21
MSFX-6 [GAPDH]	3.125 µg	86	2.7

Figure 4. The SCHAD FlexPlex™ Module MSFX-10 is designed to analyze the relative levels of SCHAD between samples. An experiment to examine levels of various enzymes of fatty acid oxidation including SCHAD is shown below. Fibroblast cell lines derived from two different patients with MCAD deficiency (MCADD) genetic disease are compared to a control fibroblast cell line (newborn human dermal fibroblast) and to a cell line with a deficiency in another enzyme of fatty acid oxidation, TFP.

Figure 5

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Figure 5. The raw signals can be compared between samples or in the example shown above the signals can be normalized to GAPDH. After normalization the patient cell line 1 has 93% reduction in MCAD and patient cell line 2 has a 92% reduction in MCAD. Both patient cell lines 1 and 2 are already described MCAD patients with the common mutation A985G. Interestingly both patients have a greater than 50% increase in mitochondrial SOD2 expression. Patient cell line 3 has a normal expression level of MCAD but a 91% decrease in TFP enzyme. Patient cell line 3 is a cell line already identified as TFP deficient at the protein level but of unknown genetic origin. Interestingly, all three patient cells lines harboring mutations in MCAD or TFP have a 50% decrease in the level of SCHAD the short chain 3-hydroxyacyl-CoA dehydrogenase.

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