Transferrin ELISA Kit

Catalog No. ABIN956352

Product Details Transferrin ELISA Kit

Target: Transferrin (TF)

Reactivity: Cow

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: The Bovine Transferrin ELISA is a highly sensitive two-site enzyme-linked immunoassay (ELISA) for the quantitative determination of transferrin in bovine biological samples.

Analytical Method: Quantitative

Characteristics: This ELISA kit can be used to measure tranferrin in serum, tissue extracts, and other biological fluids.In this assay the transferrin present in the sample reacts with the anti-Transferrin antibodies which have been adsorbed to the surface of polystyrene microtiter wells. After the removal of unbound proteins by washing, anti-Transferrin antibodies conjugated with horseradish peroxidase (HRP) are added. This HRP-conjugated antibody forms a complex with the previously bound transferrin. 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 transferrin in the sample tested, thus, the absorbance, at 450 nm, is a measure of the concentration of transferrin in the test sample. The quantity of transferrin 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-bovine transferrin 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-bovine transferrin.
7. Bovine Transferrin Calibrator: One vial containing a lyophilized Bovine Transferrin Calibrator.

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
Vortex mixer

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. Bovine Transferrin Calibrator: Add 1.0 mL of distilled or de-ionized water to the lyophilized Bovine Transferrin Calibrator and mix gently until dissolved. The calibrator is now at a concentration of 4.002 µg/mL (the reconstituted calibrator should be aliquoted and frozen if future use is intended). Bovine Transferrin Calibrators need to be prepared immediately prior to use (see the following chart). Mix well between each step. Avoid foaming.

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 transferrin 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 Bovine Transferrin Calibrator.
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-Transferrin-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-bovine transferrin 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. Bovine Transferrin Calibrator: The lyophilized Bovine Transferrin Calibrator should be stored at 4°C or frozen until reconstituted. The reconstituted calibrator should be aliquoted out and stored frozen. Avoid multiple freeze/thaw cycles. The working calibrator solutions should be prepared immediately prior to use and are stable for up to 8 hours.

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

Target Details for Transferrin

Target: Transferrin (TF)

Alternative Name: Transferrin (TF Products)

Synonyms: ltf ELISA Kit, pro1557 ELISA Kit, pro2086 ELISA Kit, mgc107777 ELISA Kit, LOC692564 ELISA Kit, TF ELISA Kit, LOC100144362 ELISA Kit, tf ELISA Kit, LTF ELISA Kit, TFEW ELISA Kit, conalbumin ELISA Kit, tf-b ELISA Kit, MGC64306 ELISA Kit, PRO1557 ELISA Kit, PRO2086 ELISA Kit, TFQTL1 ELISA Kit, AI266983 ELISA Kit, Cd176 ELISA Kit, HP ELISA Kit, Tf ELISA Kit, Tfn ELISA Kit, hpx ELISA Kit, Trf ELISA Kit, cb285 ELISA Kit, gavi ELISA Kit, id:ibd3238 ELISA Kit, id:ibd3525 ELISA Kit, sb:cb285 ELISA Kit, wu:fb57g06 ELISA Kit, wu:fb62h02 ELISA Kit, wu:fb63h10 ELISA Kit, wu:fb64h10 ELISA Kit, zgc:112154 ELISA Kit, 143958_at ELISA Kit, CG6186 ELISA Kit, Dmel\\CG6186 ELISA Kit, TSF1 ELISA Kit, anon-EST:Posey265 ELISA Kit, tsf1 ELISA Kit, Pro-TRH ELISA Kit, IL-5 ELISA Kit, STF I ELISA Kit, TRF1 ELISA Kit, sTF1 ELISA Kit, sTf ELISA Kit, tf1 ELISA Kit, transferrin ELISA Kit, serotransferrin ELISA Kit, transferrin (ovotransferrin) ELISA Kit, transferrin L homeolog ELISA Kit, melanotransferrin ELISA Kit, transferrin-a ELISA Kit, Transferrin 1 ELISA Kit, thyrotropin releasing hormone ELISA Kit, interleukin 5 ELISA Kit, TF ELISA Kit, LOC477072 ELISA Kit, tf ELISA Kit, Tf ELISA Kit, LOC100144362 ELISA Kit, tf.L ELISA Kit, LOC5575625 ELISA Kit, Trf ELISA Kit, tfa ELISA Kit, Tsf1 ELISA Kit, TRH ELISA Kit, IL5 ELISA Kit, LOC101085148 ELISA Kit, LOC100726872 ELISA Kit, trf ELISA Kit

Background: Transferring is a metal-combining protein that is reversibly bound to acid-soluble iron in plasma. It functions to transport iron to the bone marrow and to tissue storage organs such as the liver. Transferrin also participates in the regulation and control of iron absorption and protects against iron intoxication. Like haptoglobin, transferrin is returned to the circulation after unloading its iron in the reticuloendothelial system.

Pathways: Transition Metal Ion Homeostasis