Myoglobin Kit ELISA

N° du produit ABIN956102

Détail du produit Myoglobin Kit ELISA

Antigène: Myoglobin (MB)

Reactivité: Chien

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Analytical Method: Quantitative

Attributs du produit: The Dog Myoglobin ELISA is based on the principle of a solid phase enzyme-linked immunosorbent assay (ELISA). A monoclonal anti-myoglobin antibody is used for solid phase immobilization (on the microtiter wells) and polyclonal anti-myoglobin antibody conjugated to horseradish peroxidase is in the antibody-enzyme conjugate solution. The test sample is allowed to react simultaneously with the two antibodies, resulting in the myoglobin molecules being sandwiched between the solid phase and enzyme-linked antibodies. After one hour incubation at room temperature, the wells are washed to remove unbound labeled antibodies. A TMB (Tetramethyl-benzidine) Reagent is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of myoglobin is proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

Ingrédients: Anti-myoglobin-coated microtiter wells, 96 wells
Dog Myoglobin Calibrator (50 µg/mL), 50 µL
Diluent, 12 mL
Enzyme Conjugate Reagent, 11 mL
Wash Solution (20X), 50 mL
TMB Reagent (One-Step), 11 mL
Stop Solution (1N HCl), 11 mL.

Matériel non inclus: Precision pipettes
Disposable pipette tips
Distilled or de-ionized water
Vortex mixer
Absorbent paper or paper towels
Graph paper (PC graphing software is optional but recommended)
Plate shaker
Microtiter plate reader

Information d'application

Plaque: Pre-coated

Préparation de l'échantillon:

Samples may be tested undiluted or after dilution with diluent. The dilution factor should be determined empirically. On occasion a matrix effect may be observed with urine samples that may slightly increase or decrease absorbance values and we therefore strongly recommend that all urine samples within a particular study be similarly diluted. Only 20 µL of sample is required per assay (2 x 20 µL, if samples are to be tested in duplicate).
PROCEDURAL NOTES
1. Calibrators should be prepared immediately prior to use and should be used within 30 minutes of preparation.
2. Pipetting of conjugate, calibrators and samples into the microtiter plate should be completed within 10 minutes.
3. We recommend that calibrators and samples be run in duplicate.
4. It is recommended that the wells be read within 5 minutes following addition of Stop Solution.

Procédure de l'essai:

1. Ensure that all reagents are at room temperature.
   2. Secure the desired number of coated wells in the holder.
   3. Dispense 100 µL of Enzyme Conjugate Reagent into each well.
   4. Dispense 20 µL of myoglobin calibrators and samples (in duplicate) into the appropriate wells.
   5. Incubate at room temperature (18-25°C) on a plate shaker for one hour. Mix Gently (~100-150 rpm).
   6. Remove the incubation mixture either with a plate washer or by flicking plate contents into a waste container.
   7. Wash and empty the microtiter wells 5 times with 1X wash solution. This may be performed using either a plate washer (400 µL/well) or with a squirt bottle. The entire wash procedure should be performed as quickly as possible.
   8. Strike the wells sharply onto adsorbent paper or paper towels to remove all residual wash buffer.
   9. Dispense 100 µL of TMB Reagent solution into each well. Gently mix for 5 seconds.
   10. Incubate on a plate shaker at room temperature for 20 minutes. Mix Gently.
   11. Stop the reaction by adding 100 µL of Stop Solution to each well.
   12. Gently mix for 30 seconds. It is important to make sure that all the blue color changes to yellow color completely.
   13. Read absorbance at 450 nm with a microtiter plate reader within 5 minutes.

Calcul des résultats:

1. Calculate the mean absorbance values 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 graph paper, with absorbance values on the vertical or Y-axis and concentration on the horizontal or X-axis.
   3. Using the mean absorbance value for each sample, determine the corresponding concentration of myoglobin in ng/mL from the calibration curve.
   4. Multiply the derived values by the appropriate dilution factor if the test samples were diluted.
   5. Graphing software, if available, should be used.

Restrictions: For Research Use only

Stockage

Stock: -20 °C

Stockage commentaire: Store dog myoglobin calibrator at -20°C. Store the remainder of the kit at 4°C.

Détail du antigène

Antigène: Myoglobin (MB)

Autre désignation: Myoglobin (MB Produits)

Synonymes: PVALB Kit ELISA, AI325109 Kit ELISA, zgc:65819 Kit ELISA, zgc:77764 Kit ELISA, MB Kit ELISA, DKFZp468H096 Kit ELISA, myg Kit ELISA, mb Kit ELISA, MYF4 Kit ELISA, bHLHc3 Kit ELISA, myo Kit ELISA, Myoglobin Kit ELISA, myoglobin Kit ELISA, myogenin Kit ELISA, MB Kit ELISA, Mb Kit ELISA, mb Kit ELISA, myg Kit ELISA, Myog Kit ELISA

Sujet: Myoglobin is a heme protein found in both cardiac and skeletal muscle. Following myocardial necrosis associated with myocardial infarction (MI), myoglobin is one of the first markers to rise above normal levels. Studies with human subjects have shown that myoglobin increases measurably above baseline within 2-4 hours post-infarct, peaking at 9-12 hours, and returning to baseline within 24-36 hours. In the absence of skeletal muscle trauma or other factors associated with a noncardiac related increase in circulating myoglobin, myoglobin may be used as a marker for MI. Similarly, in the absence of cardiac damage, myoglobin may be used as a marker of skeletal muscle injury.

Pathways: Brown Fat Cell Differentiation