FABP3 ELISA Kit

Produktnummer ABIN956259

Dieses Affe FABP3 ELISA-Kit ist ein Colorimetric ELISA-Kit, das dafür entwickelt wurde, Affe FABP3 zu quantifizieren.

Kurzübersicht für FABP3 ELISA Kit (ABIN956259)

Target: FABP3 (Fatty Acid Binding Protein 3, Muscle and Heart (FABP3))

Reaktivität: Affe

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Applikation: ELISA

Proben: Serum

Produktdetails FABP3 ELISA Kit

Analytische Methode: Quantitative

Produktmerkmale: The Monkey H-FABP ELISA is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay uses a mouse monoclonal anti-H-FABP antibody for solid phase (microtiter wells) immobilization and a different horseradish peroxidase (HRP) conjugated mouse monoclonal anti-H-FABP antibody for detection. Calibrators and diluted samples are incubated with the HRP conjugate in the microtiter wells for 60 minutes. This results in H-FABP molecules being sandwiched between the immobilization and detection antibodies. The wells are then washed to remove unbound HRP-labeled antibodies, and TMB Reagent is added and incubated for 20 minutes at room temperature. This results in the development of a blue color. Color development is stopped by the addition of Stop Solution, changing the color to yellow, and optical density is measured spectrophotometrically at 450 nm. The concentration of H-FABP is proportional to optical density. H-FABP concentrations are determined by reference to a calibration curve.

Bestandteile: Anti-H-FABP antibody coated 96 well plate (12 strips of 8 wells)
Enzyme Conjugate Reagent, 11 mL
Reference calibrator (lyophilized)
Diluent, 50 mL
20X Wash Solution, 50 mL
TMB Reagent (One-Step), 11 mL
Stop Solution (1N HCl), 11 mL.

Benötigtes Material: Precision pipettes and tips
Distilled or de-ionized water
Polypropylene or glass tubes
Vortex mixer
Absorbent paper or paper towels
Micro-Plate incubator/shaker with an approximate mixing speed of 150 rpm
A microtiter plate reader at 450 nm wavelength, with a bandwidth of 10 nm or less and an OD range of 0-4 OD
Graph paper (PC graphing software is optional)

Anwendungsinformationen

Plattentyp: Pre-coated

Aufbereitung der Proben:

We suggest that samples initially be tested after a 5-fold dilution with the diluent provided with the kit.
1. Dispense 240 µL of 1x diluent into separate tubes.
2. Pipette and mix 60 µL of each serum sample into a tube containing 240 µL of diluent. This provides a 5-fold diluted sample.

Testdurchführung:

1. Secure the desired number of coated wells in the holder.
   2. Dispense 100 µL of calibrators and samples into the wells (we recommend that calibrators and samples be tested in duplicate).
   3. Add 100 µL of enzyme conjugate reagent into each well.
   4. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 60 minutes.
   5. Aspirate the contents of the microtiter wells and wash the wells 5 times with 1X wash solution. This may be performed using either a plate washer (400 µL/well) or with a squirt bottle. The entire wash procedure should be performed as quickly as possible.
   6. Strike the wells sharply onto adsorbent paper or paper towels to remove all residual droplets.
   7. Dispense 100 µL of TMB Reagent into each well.
   8. Gently mix on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 20 minutes.
   9. Stop the reaction by adding 100 µL of Stop Solution to each well.
   10. Gently mix. It is important to make sure that all the blue color changes to yellow.
   11. Read the optical density at 450 nm with a microtiter plate reader within 5 minutes.

Ergebnisberechnung:

1. Calculate the average absorbance values (A450) for each set of reference calibrators and samples.
   2. Construct a calibration curve by plotting the mean absorbance obtained from each reference calibrator against its concentration in ng/mL on linear graph paper, with absorbance values on the vertical or Y-axis and concentrations on the horizontal or X-axis.
   3. Using the mean absorbance value for each sample, determine the corresponding concentration of H-FABP in ng/mL from the calibration curve.
   4. Multiply the derived concentration by the dilution factor to determine the actual concentration of H-FABP in the serum sample.
   5. If available, PC graphing software may be used for the above steps.
   6. If the A450 values of samples fall outside of the calibration curve samples should be diluted appropriately and re-tested.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: The lyophilized reference calibrator must be stored at or below -20°C on receipt. The remainder of the kit should be stored at 4°C and the microtiter plate should be kept in a sealed bag with desiccant to minimize exposure to damp air. Test kits will remain stable until the expiration date provided that the components are stored as described above.

Haltbarkeit: The expiry date is stated on the label.

Details zu FABP3

Target: FABP3 (Fatty Acid Binding Protein 3, Muscle and Heart (FABP3))

Andere Bezeichnung: H-FABP

Hintergrund: Fatty acid-binding proteins (FABP) are cytosolic proteins of about 15 kDa. They bind long chain fatty acids and play an important role in fatty acid metabolism. Heart, liver and intestinal FABP isoforms exist. Heart has a high content of FABP (10-20 mol % of cytoplasmic proteins) and heart FABP (H-FABP) has proved to be a sensitive biomarker of myocardial necrosis in humans. H-FABP is rapidly released into the circulation from damaged cardiac muscle. In humans serum levels increase significantly within 1-4 hours of muscle injury and return to normal within 12 to 24 hours. Because H-FABP is also expressed in skeletal muscle, it is necessary to exclude or control for skeletal muscle inquiry before ascribing H-FABP elevations to cardiac injury. However, in the absence of cardiac injury H-FABP is a useful biomarker of skeletal muscle injury. Validation studies at revealed basal monkey H-FABP levels of ~ 20 ng/mL with levels in excess of 400 ng/mL in animals with muscle injury.

Pathways: Monocarboxylic Acid Catabolic Process