FABP1 ELISA Kit

Catalog No. ABIN956112

The Rat FABP1 ELISA Kit (ABIN956112) is a Colorimetric ELISA Kit designed to quantify Rat FABP1.

Quick Overview for FABP1 ELISA Kit (ABIN956112)

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

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Serum

Product Details FABP1 ELISA Kit

Analytical Method: Quantitative

Specificity: Cross-reactivity with intestinal FABP has not been investigated.

Characteristics: The Rat L-FABP ELISA is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay uses affinity purified anti-rat L-FABP antibodies for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated anti-rat L-FABP antibodies for detection. The test sample is diluted and incubated with conjugate in the microtiter wells for 60 minutes. This results in L-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 L-FABP is proportional to the optical density of the test sample and the actual value is derived by reference to a calibration curve.

Components: Anti-rat L-FABP antibody coated microtiter plate with 96 wells (provided as 12 detachable strips of 8)
Enzyme Conjugate Reagent, 11 mL
Rat L-FABP Calibrator (lyophilized), containing 2 µg/mL rat L-FABP when reconstituted as detailed on the vial label
Diluent, 25 mL
Wash Solution (20X), 50 mL
TMB Reagent (One-Step), 11 mL
Stop Solution (1N HCl), 11 mL.

Material not included: 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)

Application Details

Plate: Pre-coated

Sample Preparation:

Serum samples may be tested undiluted or after dilution with diluent. The optimal dilution factor should be determined empirically.

Assay Procedure:

1. Secure the desired number of coated wells in the holder.
   2. Dispense 100 µL of calibrators and diluted samples into the wells (we recommend that 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. Wash and empty the microtiter wells 5 times with 1X wash solution. Optimal results are obtained if a plate washer (400 µL/well) is used. However, if a plate washer is not available a squirt bottle may be used to manually wash the wells. 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.

Calculation of Results:

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 concentration on the horizontal or X-axis.
   3. Using the mean absorbance value for each sample, determine the corresponding concentration of L-FABP in ng/mL from the calibration curve.
   4. Multiply the derived concentration by the dilution factor to determine the actual concentration of L-FABP in the serum/plasma sample.
   5. If available, PC graphing software may be used for the above steps.
   6. If the OD450 values of samples fall outside of the calibration curve samples should be diluted appropriately and re-tested.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: The unused 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.

Expiry Date: The expiry date is stated on the label.

Target Details for FABP1

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

Alternative Name: L-FABP

Background: Liver Fatty Acid Binding Protein (L-FABP) has a molecular weight of ~ 14 kDa and constitutes 2-5% of liver cytosolic protein. Immunologically distinct intestinal and cardiac FABP isoforms also exist. All isoforms serve a role in fatty acid transport and metabolism. L-FABP is expressed primarily in the liver but relatively high levels are found in intestinal tissue and it is expressed in other tissues also. L-FABP has recently been identified as a possible biomarker for lung, kidney and liver disease. The L-FABP kit manufactured by does not recognize rat cardiac FABP.

Pathways: Chromatin Binding, Regulation of Lipid Metabolism by PPARalpha