Ferritin ELISA Kit

Catalog No. ABIN956076

Product Details Ferritin ELISA Kit

Target: Ferritin (FE)

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: The Mouse Ferritin ELISA is a highly sensitive two-site enzyme-linked immunoassay (ELISA) for the quantitative determination of Ferritin in mouse biological samples.

Analytical Method: Quantitative

Characteristics: In this assay the ferritin present in the sample reacts with the anti-ferritin antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-ferritin antibodies conjugated with horseradish peroxidase (HRP) are added. This HRP-conjugated antibody forms a complex with the previously bound ferritin. 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 ferritin in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of ferritin in the test sample. The quantity of ferritin in the test sample can be interpolated from the calibration curve constructed from the calibrators, and corrected for sample dilution.

Components: 1. Diluent Concentrate: One bottle containing 50 mL of a 5X 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-mouse ferritin antibody conjugated with HRP in 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. Wells are coated with affinity-purified anti-mouse ferritin.
7. Mouse Ferritin Calibrator: One vial containing 1.2 mL of Mouse Ferritin Calibrator.
8. Positive Control: One vial containing 50 µL of serum with 0.1% sodium azide. See the Control Certificate for the concentration.

Material not included: 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

Application Details

Plate: Pre-coated

Reagent Preparation:

1. Diluent Concentrate: The Diluent solution supplied is a 5X concentrate and must be diluted 1:5 with distilled or de-ionized water.
   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. Mouse Ferritin Calibrator The Mouse Ferritin Calibrator should be aliquoted out and stored frozen. It is at a concentration of 1.117 µg/mL. Mouse Ferritin Calibrators need to be prepared immediately before use. Mix well between each step. Avoid foaming.
   8. Positive Control: The concentration and recommended dilution are provided on the Control Certificate. Before use, briefly centrifuge the Positive Control to allow all of the liquid to collect in the bottom of the vial.

Calculation of Results:

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 ferritin 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.

Restrictions: For Research Use only

Handling

Storage: 4 °C

Storage Comment: 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 Mouse Ferritin Calibrator and Positive Control.
2. Diluent: The 5X Diluent 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 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-Ferritin-HRP conjugate should be stored at 4°C and diluted immediately prior to use. The working conjugate solution is stable for up to 1 hour when stored in the dark.
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-mouse ferritin 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. Mouse Ferritin Calibrator: Long Term Storage : Upon receipt, aliquot the calibrator and store them frozen. They will be stable until expiration date.

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

Target Details for Ferritin

Target: Ferritin (FE)

Alternative Name: Ferritin (FE Products)

Synonyms: 1HCH ELISA Kit, BEST:LD36673 ELISA Kit, CG2216 ELISA Kit, Dmel\\CG2216 ELISA Kit, FER1 ELISA Kit, FER1HCH ELISA Kit, Fer-L ELISA Kit, Fer1 ELISA Kit, Fer1HC ELISA Kit, FerHCH ELISA Kit, HCH ELISA Kit, T21 ELISA Kit, T9 ELISA Kit, fer1 ELISA Kit, fer1hch ELISA Kit, l(3)00451 ELISA Kit, l(3)j10B4 ELISA Kit, SFerH-1 ELISA Kit, SOF-35 ELISA Kit, SOF-5L ELISA Kit, NV13011 ELISA Kit, ACP1 ELISA Kit, F8A5.24 ELISA Kit, F8A5_24 ELISA Kit, Ferritin 1 heavy chain homologue ELISA Kit, ferritin light chain ELISA Kit, ferritin ELISA Kit, chromosome 26 open reading frame, human C10orf131 ELISA Kit, hyaluronan and proteoglycan link protein 2 ELISA Kit, NAD(P)-linked oxidoreductase superfamily protein ELISA Kit, Fer1HCH ELISA Kit, H-1 ELISA Kit, YPDSF_1340 ELISA Kit, ftnA ELISA Kit, Ft ELISA Kit, C26H10orf131 ELISA Kit, HAPLN2 ELISA Kit, AT1G60730 ELISA Kit

Background: Ferritin is a water-soluble, iron storage protein. Serum ferritin level is said to be useful for studying iron deficiency anemia, metabolism disorder and malignant tumor. Ferritin may also be an acute-phase protein and is often elevated in the course of disease.

Pathways: Transition Metal Ion Homeostasis