FABP1 ELISA Kit

Produktnummer ABIN956355

Das Ratte FABP1 ELISA Kit (ABIN956355) ist ein Colorimetric ELISA Kit zur Detektion von Ratte FABP1.

Kurzübersicht für FABP1 ELISA Kit (ABIN956355)

Target: FABP1 (Fatty Acid Binding Protein 1, Liver (FABP1))

Reaktivität: Ratte

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Applikation: ELISA

Produktdetails FABP1 ELISA Kit

Verwendungszweck: The Rat L-FABP ELISA is a highly sensitive two-site enzyme-linked immunoassay (ELISA) for the quantitative determination of L-FABP in rat biological samples.

Analytische Methode: Quantitative

Produktmerkmale: In this assay the L-FABP present in the sample reacts with the anti-L-FABP antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-L-FABP antibodies conjugated with horseradish peroxidase (HRP) are added. This HRP-conjugated antibody forms a complex with the previously bound L-FABP. Following another washing step, the enzyme bound to the immunosorbent is assayed by the addition of a chromogenic substrate, 3,3',5,5'- tetramethylbenzidine (TMB). The quantity of bound enzyme is proportional to the concentration of L-FABP in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of L-FABP in the test sample. The quantity of L-FABP in the test sample can be interpolated from the calibration curve constructed from the calibrators, and corrected for sample dilution.

Bestandteile: 1. Diluent Concentrate: One bottle containing 60 mL of a 1X concentrated diluent running buffer.
2. Wash Solution Concentrate: One bottle containing 50 mL of a 20X concentrated wash solution.
3. Enzyme-Antibody Conjugate Concentrate: One vial containing 150 µL of a 100X concentrated affinity-purified anti-rat L-FABP antibody conjugated with HRP in a stabilizing buffer.
4. TMB Substrate Solution: One vial containing 12 mL of TMB and hydrogen peroxide in citric acid buffer at pH 3.3.
5. Stop Solution: One vial containing 12 mL of 0.3 M sulfuric acid. WARNING: Avoid contact with skin.
6. Microtiter Plate: Twelve removable eight-well micro strips in well holder frame. Each well is coated with affinity-purified anti-rat L- FABP.
7. Rat L-FABP Calibrator: One vial containing Rat L-FABP Calibrator.

Benötigtes Material: Precision pipettes (2 µL to 200 µL) for making and dispensing dilutions
Test tubes
Microplate washer/aspirator
Distilled or de-ionized H2O Microplate reader
Assorted glassware for the preparation of reagents and buffer solutions
Timer
Vortex mixer

Anwendungsinformationen

Plattentyp: Pre-coated

Aufbereitung der Reagenzien:

1. Diluent Concentrate: The Diluent solution supplied is a 1X concentrate. No dilution is required.
   2. Wash Solution Concentrate: The Wash Solution supplied is a 20X concentrate and must be diluted 1:20 with distilled or de-ionized water. Crystal formation in the concentrate is not uncommon when storage temperatures are low. Warming of the concentrate to 30-35°C before dilution can dissolve crystals.
   3. Enzyme-Antibody Conjugate Concentrate: Calculate the required amount of working conjugate solution for each microtiter plate test strip by adding 10 µL Enzyme-Antibody Conjugate to 990 µL of 1X Diluent for each test strip to be used for testing. Mix uniformly, but gently. Avoid foaming.
   4. TMB Substrate Solution: Ready to use as supplied.
   5. Stop Solution: Ready to use as supplied.
   6. Microtiter Plate: Ready to use as supplied. Unseal Microtiter Pouch and remove plate from pouch. Remove all strips and wells that will not be used in the assay and place back in pouch and re-seal along with desiccant.
   7. Rat L-FABP Calibrator The Rat L-FABP Calibrator should be aliquoted and stored frozen. It is at a concentration of 23.08 µg/mL and needs to be diluted in 1X diluent according to the chart below for each run. Rat L-FABP Calibrators need to be prepared immediately prior to use (see the following chart). Mix well between each step. Avoid foaming.

Ergebnisberechnung:

1. Subtract the average background value from the test values for each sample.
   2. Using the results observed for the calibrators construct a calibration curve. The appropriate curve fit is that of a four- parameter logistics curve. A second order polynomial (quadratic) or other curve fits may also be used.
   3. Interpolate test sample values from calibration curve. Correct for sample dilution factor to arrive at L-FABP concentration in original sample.
   
   1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the information contained in the package insert instructions and with adherence to good laboratory practice.
   2. Factors that might affect the performance of the assay include proper instrument function, cleanliness of glassware, quality of distilled or de-ionized water, and accuracy of reagent and sample pipettings, washing technique, incubation time or temperature.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C

Informationen zur Lagerung: 1. Complete Kit: The expiration date for the kit is stated on the outer label. The recommended storage temperature is 4°C. Note: See long term storage recommendations below for the Rat L-FABP Calibrator.
2. Diluent: The 1X Diluent Concentrate is stable until the expiration date. The solution should be stored at 4°C.
3. Wash Solution: The 20X Wash Solution Concentrate is stable until the expiration date. The 1X working solution is stable for at least one week from the date of preparation. Both solutions can be stored at room temperature (RT, 16-25°C) or at 4°C.
4. Enzyme-Antibody Conjugate: Undiluted anti-L-FABP-HRP conjugate should be stored at 4°C and diluted immediately prior to use. The working conjugate solution is stable for up to 8 hours.
5. TMB Substrate Solution: The TMB Substrate Solution should be stored at 4°C and is stable until the expiration date.
6. Stop Solution: The Stop Solution should be stored at 4°C and is stable until the expiration date.
7. Microtiter Plate: Anti-rat L-FABP coated wells are stable until the expiration date and should be stored at 4°C in the sealed foil pouch with a desiccant pack.
8. Rat L-FABP Calibrator: Long Term Storage: Upon receipt, aliquot the calibrator and store frozen. They will be stable until expiration date. Short term. Storage : The calibrator is stable for up to 14 days at 4°C. The working calibrator solutions should be prepared immediately prior to use.

Haltbarkeit: The expiry date is stated on the label.

Details zu FABP1

Target: FABP1 (Fatty Acid Binding Protein 1, Liver (FABP1))

Andere Bezeichnung: L-FABP

Pathways: Chromatin Binding, Regulation of Lipid Metabolism by PPARalpha