IgE Kit ELISA

N° du produit ABIN956263

Kit ELISA IgE Singe, Colorimetric test pour la quantification de Singe IgE.

Aperçu rapide pour IgE Kit ELISA (ABIN956263)

Antigène: IgE

Reactivité: Singe

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Type d'échantillon: Plasma, Serum

Détail du produit IgE Kit ELISA

Analytical Method: Quantitative

Specificité: Cross-reactivity with IgE from other species has not been investigated. IgE is the least abundant immunoglobulin in serum, typically present at levels of 1 µg/mL per mL or lower. It is involved in allergic reactions. It binds to Fc receptors on basophils and mast cells. Subsequent binding of antigen to IgE triggers release of histamine and other vasoactive amines. IgE levels are elevated in asthma, eczema, rhinitis and parasitic infections. IgE is useful as a serum biomarker in such conditions.

Attributs du produit: The monkey IgE ELISA kit is designed for measurement of IgE in old world monkey serum or plasma. The assay uses a mouse monoclonal anti-monkey IgE for solid phase (microtiter wells) immobilization and a different horseradish peroxidase (HRP) conjugated mouse monoclonal anti-monkey IgE antibody for detection. The kit will also recognize human IgE but this kit is not intended for research or diagnostic testing of human samples.Test samples are diluted and incubated in the microtiter wells for 45 minutes alongside prepared monkey IgE calibrators. The microtiter wells are subsequently washed and HRP conjugate is added and incubated for 45 minutes. IgE molecules are thus 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 IgE is proportional to the optical density of the test sample and is derived from a calibration curve.

Ingrédients: Anti monkey IgE coated 96-well plate (12 strips of 8 wells)
Reference calibrator (lyophilized). Store at -20°C.
10X Diluent, 25 mL
HRP Conjugate Reagent, 11 mL
20X Wash Solution, 50 mL
TMB Reagent (One-step), 11 mL
Stop Solution (1N HCl), 11 mL.

Matériel non inclus: Precision pipettes and tips
Distilled or de-ionized water
Vortex mixer
Absorbent paper or paper towels
Graph paper (PC graphing software is optional)
Polypropylene or glass tubes
Plate reader with an optical density range of 0-4 at 450 nm.
Micro-Plate incubator/shaker mixing speed of ~150 rpm
Plate washer

Information d'application

Plaque: Pre-coated

Préparation de l'échantillon:

General Note: We found IgE to present in a panel of Rhesus and Cynomolgus monkey serum samples at concentrations of ~0.2 to 10 µg/mL, with most samples in the 0.2 to 1 µg/mL range. In order to obtain values within range of the calibration curve, we suggest that samples initially be diluted 200 fold using the following procedure for each sample to be tested:
1. Dispense 248.75 µL of diluent into a microcentrifuge tube.
2. Pipette and mix 1.25 µL of the serum/plasma sample into the 248.75 µL of diluent. This provides a 200 fold diluted sample. Do not use sample dilutions less than 40-fold (i.e., 20 fold). At dilutions less than 40-fold serum factors may interfere with the assay.

Procédure de l'essai:

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. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (25°C) for 45 minutes.
   4. Aspirate the contents of the microtiter wells and wash the wells 5 times with 1x wash solution using a plate washer (400 µL/well). The entire wash procedure should be performed as quickly as possible.
   5. Strike the wells sharply onto absorbent paper or paper towels to remove all residual wash buffer.
   6. Add 100 µL of enzyme conjugate reagent into each well.
   7. Incubate on an orbital micro-plate shaker at 100-150 rpm at room temperature (25°C) for 45 minutes.
   8. Wash as detailed in 4 to 5 above.
   9. Dispense 100 µL of TMB Reagent into each well.
   10. Gently mix on an orbital micro-plate shaker at 100-150 rpm at room temperature (18-25°C) for 20 minutes.
   11. Stop the reaction by adding 100 µL of Stop Solution to each well.
   12. Gently mix. It is important to make sure that all the blue color changes to yellow.
   13. Read the optical density at 450 nm with a microtiter plate reader within 5 minutes.

Calcul des résultats:

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

Restrictions: For Research Use only

Stockage

Stock: 4 °C/-20 °C

Stockage commentaire: The lyophilized reference calibrator should be stored in a freezer at or below -20°C when the kit is received. The remainder of the kit should be stored in a refrigerator at 4°C and must not be frozen. The test kit will remain stable until the expiration date provided that the components are stored as described above. The microtiter plate should be kept in a sealed bag with desiccant to minimize exposure to damp air.

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

Détail du antigène

Antigène: IgE

Classe de substances: Antibody