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Succinate Assay Kit (BA0149)

SKU:
BA0149
Product Type:
Assay
Instrument:
Microplate Reader
Sample Type:
Cells, Tissue
Research Area:
TCA Cycle
Diabetes & Obesity
Food Safety & Analysis
Plant & Environmental Stress
Clinical Chemistry
$1,511
Frequently bought together:

Description

ELISA Kit Technical ManualMSDS

About Succinate:

SUCCINATE, or succinic acid, can be found in all plants and animal tissues. It is an intermediate in the citric acid cycle and plays an important role in intracellular energy generation. Succinate is widely used as a flavoring agent in the food, beverage, and pharmaceutical industries due to its low toxicity.

How our Succinate Assay Kit Works:

Assay Genie's succinate assay provides a simple, one step assay for measuring succinate. In this assay succinate is converted to pyruvate which reacts with specific reagents and dye to form a colored product. The color intensity at 570 nm or fluorescence at Ex/Em = 530/585 nm of the reaction product is directly proportional to succinate concentration in the sample.

Key Features

  • Fast and sensitive. Use 20 uL sample. Linear detection range 10 to 400 uM for colorimetric assays and 2 to 40 uM for fluorimetric assays.
  • Convenient. The procedure involves adding a single working reagent, and reading the absorbance or fluorescence after 30 minutes. Room temperature assay. No 37°C heater is needed.
  • High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Succinate Assay Kit Information:

Features
Applications:
For quantitative determination of succinate (succinic acid) in food, beverage, agricultural products, and other biological samples.
Kit Includes:
  • Assay Buffer: 10 mL
  • Enzyme Mix: 120 µL
  • Co substrate: 120 µL
  • PEP: Dried Dye Reagent: 120 µL
  • Standard: 500 µL
  • 20 mM Succinate
Kit Requires:
Pipetting devices, centrifuge tubes, clear flat-bottom 96-well plates, and plate or cuvette reader.
Method of Detection:
  • Colorimetric: OD570nm
  • Fluorimetric: FL530/585nm
Detection Limit:
  • Colorimetric assay: 10 µM
  • Fluorimetric assay: 2 µM
Samples:

Food, beverage, agricultural products, and other biological samples.
Species:
All
Protocol Length:
30 min
Size:
100 tests
Storage:
Store all components at -20°C upon receiving
Shelf Life:
6 months

Succinate Assay - Colorimetric Procedure:

Step Procedure

1.

Internal standard is required for colorimetric assay. Each sample requires two separate reactions: 1) Sample plus internal standard and 2) Sample alone. In addition, each assay plate requires a water blank well. Add 20 µL of each sample to two separate wells. Also, add 20 µL dH2O to a separate well. For the internal standard, prepare 400 µL 1 mM succinate standard by mixing 20 µL 20 mM standard with 380 µL dH2O. Add 5 µL 1 mM standard to the sample plus internal standard wells. Add 5 µL dH2O to the sample alone and water wells.

2.

Prepare sufficient Working Reagent (WR) for all wells by mixing, for each well, 85 µL Assay Buffer, 1 µL Enzyme Mix, 1 µL Cosubstrate, 1 µL PEP and 1 µL dye reagent. Fresh reconstitution of the WR is recommended. Add 80 µL WR to each well. Tap plate to mix. Incubate for 30 min at room temperature.

3.

Read optical density at 570nm (550-585nm).

Succinate Assay - Fluorimetric Procedure:

Step Procedure

1.

Prepare a 40 µM Standard Premix by mixing 20 µL of 1 mM succinate (see colorimetric internal standard procedure) with 480 µL dH2O. Dilute Standard in distilled water as follows:
Number Premix + H2O Vol (µL) Succinate (µM)

1.

100 µL + 0 µL

100

40

2.

60 µL + 40 µL

100

24

3.

30 µL + 70 µL

100

12

4.

0 µL + 100 µL

100

0

Transfer 20 µL standards and 20 µL samples into separate wells of a black 96-well plate.

2.

Add 80 µL Working Reagent (see Colorimetric Procedure). Tap plate to mix.

3.

Incubate 30 min at room temperature and read fluorescence at ex/em = 530/585 nm.

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