Haptoglobin Kit ELISA

N° du produit ABIN955897

Détail du produit Haptoglobin Kit ELISA

Antigène: Haptoglobin (HP)

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Fonction: The Mouse Haptoglobin ELISA is a highly sensitive two-site enzyme-linked immunoassay (ELISA) for the quantitative determination of haptoglobin in mouse serum or plasma.

Analytical Method: Quantitative

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

Ingrédients: 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 haptoglobin 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 haptoglobin.
7. Mouse Haptoglobin Calibrator: One vial containing a lyophilized Mouse Haptoglobin Calibrator.
8. Positive Control: One vial containing 50 µL of serum with 0.1% sodium azide. See the Control Certificate for the concentration.

Matériel non inclus: 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

Information d'application

Plaque: Pre-coated

Préparation des réactifs:

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 Haptoglobin Calibrator: Add 1.0 mL of distilled or de-ionized water to the lyophilized Mouse Haptoglobin Calibrator and mix gently until dissolved. The calibrator is now at a concentration of 12.09 µg/mL (the reconstituted calibrator should be aliquoted and frozen if future use is intended). Mouse Haptoglobin Calibrators need to be prepared immediately prior to use (see the following chart). 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.

Calcul des résultats:

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 haptoglobin 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

Stockage

Stock: 4 °C

Stockage commentaire: 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 Haptoglobin 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-Hp-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-mouse haptoglobin 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 Haptoglobin Calibrator: The lyophilized Mouse Haptoglobin Calibrator should be stored at 4°C or frozen until reconstituted. The reconstituted calibrator should be aliquoted 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.
9. Positive Control: For storage longer than 7 days keep frozen until the expiration date. Storage less than 7 days can be at 4°C. Avoid multiple freeze/thaw cycles.

Date de péremption: The expiry date is stated on the label.

Détail du antigène

Antigène: Haptoglobin (HP)

Autre désignation: Haptoglobin (HP Produits)

Synonymes: HP Kit ELISA, wu:fb64e01 Kit ELISA, BP Kit ELISA, HP2ALPHA2 Kit ELISA, HPA1S Kit ELISA, HP-1 Kit ELISA, HPR Kit ELISA, Zonulin Kit ELISA, haptoglobin Kit ELISA, haptoglobin-like Kit ELISA, HP Kit ELISA, hp Kit ELISA, Hp Kit ELISA, LOC479668 Kit ELISA, LOC101102413 Kit ELISA

Sujet: Acute phase proteins are plasma proteins which increase in concentration following infection, inflammation or trauma. The first acute phase protein to be recognized was discovered in humans by Tillet and Frances in 1930. Haptoglobin (Hp) is a heterogeneous plasma protein mostly synthesized in the liver. The haptoglobin monomer consists of two heavy chains, beta chains (40 kDa) and two light chains, alpha 1 chain (9 kDa) and alpha 2 chain (16 kDa) that are linked by disulfide bonds. The three major haptoglobin types are, Hp1-1 which is monomeric at 98 kDa, Hp1-2 is polymeric at about 200 kDa and Hp2-2 at about 400 kDa. The haptoglobin levels in serum rise quickly following acute tissue damage within 24 to 48 hours and fall quickly once the stimulus is removed. In fact, haptoglobin levels are decreased in hemolytic anemia. Haptoglobin has a high affinity for hemoglobin (Hb) and its function appears to be to prevent loss of Hb in urine which would lead to loss of iron. Investigations over the past few years have shown that quantification of haptoglobin in plasma and serum can provide valuable information in the detection, prognosis and monitoring of disease not only in humans but in companion animals and farm herds as well.

Pathways: Transition Metal Ion Homeostasis