ImmunoDiagnostics Human Fatty-Acid Binding Protein 4 ELISA kit

FABP4 Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total.  The wells are coated with a mouse monoclonal anti- human FABP4 antibody.  The microplate is sealed in a foil bag.  Any unused strips should be returned to the bag and resealed for future use.

Detection Antibody Solution: Concentrate (100x conc). One vial containing 0.12 mL of a biotin conjugated polyclonal anti- human FABP4.  Detection antibody should be diluted with only the 1X Assay Buffer needed.

Assay Buffer: Concentrate (5x conc).  One vial containing 20 mL of buffer for dilution of the detection antibody.  If precipitates are observed in the concentrated buffer, warm at 37^oC until the precipitates disappear.  Dilute 20 mL of 5x Assay Buffer with 80 mL diH₂O to make a 1x concentration prior to use.

Human FABP4 Standard: 25 ng of recombinant human FABP4 in a buffered protein base, lyophilized.  Reconstitute with 1 mL 1x Assay Buffer to prepare 25 ng/mL standard stock.  Prepare a serial dilution series with 1x Assay Buffer.  The reconstituted standard stock can be aliquoted and stored at -20^oC for one month.  Avoid repeated freeze/thaw cycles.

Wash Buffer: Concentrate (10x conc).  One vial containing 50 mL of 10x wash buffer.  If precipitates are observed in the 10x solution, warm at 37^oC until the precipitates disappear. Prepare 1x Wash Buffer my mixing 50 mL of 10x Wash Buffer with 450 mL diH₂O.

STP-HRP Solution: Concentrate (200x conc).  One vial containing 0.06 mL of 200x STP-HRP solution.  Briefly spin the 100x vial and dilute the desired amount of 200x STP-HRP solution 1:200 with 1x Assay Buffer to prepare the 1x solution.  100 μL of 1x solution are required per well.  Immediately return unused concentrate to 2-8^oC storage.

Substrate Solution: One bottle containing 12 mL of TMB (3,3’,5,5’-Tetramethylbenzidine) substrate.  Ready-to-use.  Do not expose to light.

Stop Solution: One vial containing 12 mL of ready-to-use stop solution.

The FABP4 ELISA is a quantitative sandwich enzyme immunoassay. Standards, diluted samples, and desired controls are pipetted into the wells pre-coated with solid-phase capture antibody. After washing away any unbound substances, a biotin-labelled detection antibody, specific for human FABP4, is added to the wells, and any unbound antibody is washed out. Streptavidin-HRP conjugate is then added and a TMB solution is added to develop color in proportion to the amount of human FABP4 initially bound. The color development is stopped by addition of an acidifying solution and the optical density is determined. By plotting a standard curve versus measured absorbance, the amount of antigen in the sample can be calculated. The concentration of the antigen is expressed in ng/mL.

The kit contains calibrators and can be used in any laboratory equipped with a plate reader. The kit is optimized for the quantitative determination of human FABP4 concentrations in serum plasma, and cell culture supernatant samples. The test can provide results within 3 hours.

Example only! Do not use for calculations

4 Parameter Curve

Log-log Regression Curve

Fatty-acid binding protein 4 (FABP4), also called adipocyte Protein 2 (aP2), is a novel adipocyte-expressed factor which accounts for ~6% of total cellular proteins. Mice with targeted disruption of FAPB4 accompanying FABP5 almost completely protect against diet-induced obesity, insulin resistance, dyslipidemia, type 2 diabetes, and fatty liver disease. Secreted FABP4 correlated positively with obesity, non-alcoholic fatty liver disease (NAFLD), insulin resistance, and Type 2 Diabetes Miletus.

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