1. Catalogs
  2. Calbiotech
  3. Myoglobin ELISA

Myoglobin ELISA

Myoglobin ELISA

Myoglobin ELISA

Product catalog summary
Intended Use
The Calbiotech Inc. Myoglobin ELISA Kit is designed for the quantitative determination of myoglobin in human serum or plasma.
Summary and Explanation
Myoglobin is a heme protein found in cardiac and skeletal muscle. It is released into the serum following muscle damage, such as myocardial infarction (MI). Myoglobin levels rise within 2-4 hours post-infarct, peak at 9-12 hours, and return to baseline within 24-36 hours. It serves as an early marker for MI, with high negative predictive values reported for ruling out myocardial infarction.
Principle of the Test
The Myoglobin ELISA test uses a solid phase enzyme-linked immunosorbent assay. It employs a monoclonal antibody against myoglobin for solid phase immobilization and a goat anti-myoglobin antibody in the enzyme conjugate solution. The test sample reacts with these antibodies, forming a sandwich complex. After incubation and washing, a TMB reagent is added, developing a color proportional to myoglobin concentration, measured at 450 nm.
Materials Provided
  • Microwell coated with murine monoclonal anti-myoglobin
  • Reference Standard Set
  • Sample Diluent
  • Enzyme Conjugate Reagent
  • TMB Reagent
  • Stop Solution
  • Wash Concentrate
Materials Not Provided
  • Distilled or deionized water
  • Precision pipettes
  • Disposable pipette tips
  • ELISA reader
  • Absorbance paper
  • Graph paper
Storage and Stability
Store the kit at 2-8°C. Keep microwells sealed with desiccants. Reagents are stable until expiration. Avoid exposure to heat, sun, or strong light.
Warnings and Precautions
  • For research use only, not for diagnostic procedures.
  • Handle reagents at Biosafety Level 2 due to potential biohazardous materials.
  • Adhere strictly to the test protocol for optimal results.
  • Do not mix components from different kit lots.
Reagent Preparation
Bring all reagents to room temperature before use. Dilute patient and control serum 10-fold. Prepare 1X Wash buffer by diluting the concentrate with distilled water.
Assay Procedure
  1. Secure coated wells in the holder.
  2. Dispense 20 µl of standards, diluted specimens, and controls into wells.
  3. Add 200 µl of Enzyme Conjugate Reagent to each well and mix.
  4. Incubate at room temperature for 45 minutes.
  5. Wash wells three times with 1X wash buffer.
  6. Add 100 µl of TMB Reagent to each well and incubate for 20 minutes.
  7. Stop the reaction with 100 µl of Stop Solution and read absorbance at 450 nm.
Standard Curve
The standard curve provides absorbance values at 450 nm for myoglobin concentrations ranging from 0 to 1000 ng/ml.
See more

Catalog excerpts

Myoglobin ELISA-1

Juronen, E.L., Viikmaa, M.H., and Mikelsarr, A.V.N.: Rapid, simple and sensitive antigen capture ELISA for the quantitation of myoglobin using monoclonal antibodies, J. Imm. Meth.: 111, 109; (1988). Chapelle JP. et al.: Serum myoglobin determinations in the assessment of acute myocardial infarction. Eur. Heart Journal, 3:122, (1982). Cairns, J.P., et.al.: Usefulness of serial determinations of myoglobin and creatine kinase in serum compared for assessment of acute myocardial infarction, Clin. Chem. News, 29: 469, (1983). Silva, D.P., et.al.: Development and application of antibodies to human cardiac myoglobin in rapid fluolrescence immunoassay, Clin. Chem., 37: 1356, (1991). Ellis AK.: Patters of myoglobin release after reperfusion of injured myocardium. Clin. Chem., 72:639, (1985). Mair J. et al.: Rapid diagnosis of myocardial infarction by immunoturbidimetric myoglobin measurement (letter). Lancet, ;337:1343, (1991). Chapelle, J.P.: Myoglobin. Clin. Chem. News, 17:22, (1991). Hamfelt, A., et. al.: Use of biochemical tests for myocardial infarction in the county of Vasternorrland, a clinical chemistry routine for the diagnosis of myocardial infarction. Scand. J. Clin. Lab. Invest. Suppl., 200:20, (1990). Tucker, J.F., et.al.: Early diagnostic efficiency of cardiac troponin I and Troponin T for acute myocardial infarction, Academic Emergency Medicine: 4(1): 13-21; (1997). de Winter, R.J., et.al.: Value of myoglobin, troponin T and CK-Mbmass in ruling out an acute myocardial infarction in the emergency room, Circulation: 92(12): 3401-7; (1995). Montague, C., Kircher, T.: Myoglobin in early evaluation of acute chest pain, Amer. J. Clin. Path.: 104(4): 472-6; (1995). Tucker, J.F., et.al., Value of serial myoglobin levels in the early diagnosis of patients admitted for acute myocardial infarction, Annals of Emergency Medicine: 24(4): 704-8; (1994). Roxin, L.E., et.al.: The value of serum myoglobin determinations in the early diagnosis of acute myocardial infarction, Acta Medica Scand.: 215(5): 417-25; (1984). Sylven, C., Bendz, R.: Myoglobin, creatine kinase and its isoenzyme MB in serum after acute myocardial infarction, Eur. J. Cardiol.: 8(4-5): 515-21; (1978). Norregaard-Hansen, K., et. al.: Early observations of S-myoglobin in the diagnosis of acute myocardial infarction. The influence of discrimination limit, analytical quality, patient’s sex, and prevalence of disease. Scand. J. Clin. Lab. Invest., 46:561-569, (1986). Engvall, E., “Methods in Enzymology”, Volume 70, VanVunakis H. and Langone, J.J. (eds.), Academic Press, New York, NY, 419-492, (1980). Uotila, M., Ruouslahti, E. And Engvall, E., J. Immunol. Methods, 42, 11-15, (1981). U.S. Department of Labor, Occupational Safety and Health Administration, 29 CFR Part 1910.1030. Occupational Exposure ot Bloodborne Pathogens; Final Rule. Federal Register; 56(235):64175, (1991). USA Center for Disease Control/National Institute of Health Manual, “Biosafety in Microbiological and Biomedical Laboratories”, (1984). National Committee for Clinical Laboratory Standards. Protection of Laboratory Workers from Instrument Biohazards and Infectious Disease Transmitted by Blood, Body Fluids, and Tissue: Approved Guideline. NCCLS Document M29-A, (1997). Clinical Guide to Laboratory Tests. N.W. Tietz, Ed., 3rd Edition, W.B. Saunders, Co., p. 482, (1995). 2017-04-17 Cat#: MG017C (96 Tests) For Order and Inquiries, please contact Calbiotech Inc., 1935 Cordell Ct., El Cajon, CA 92020 Tel (619) 660-6162, Fax (619) 660-6970, www.calbiotech.com Myoglobin ELISA Catalog No.: MG017C (96 Tests) INTENDED USE The Calbiotech Inc. (CBI) Myoglobin ELISA Kit is intended for the quantitative determination of Myoglobin in human serum or plasma. SUMMARY AND EXPLANATION Myoglobin, a heme protein with a molecular weight of approximately 17,500 Daltons is found in both cardiac and skeletal muscle. Damage to either type of muscle following conditions such as trauma, ischemia, and diseases that cause myopathy, is associated with the release of myoglobin into serum. Specifically, following cardiac necrosis associated with myocardial infarction (MI), myoglobin is one of the first markers to rise above normal levels. Myoglobin levels increase 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 non-cardiac related increase in circulating myoglobin, its levels have been used as an early marker for myocardial infarct. A number of reports suggest using the measurement of myoglobin as a diagnostic aid in ruling out myocardial infarction with negative predictive values of up to 100% reported at certain time periods after the onset of symptoms.915 Unlike the other cardiac enzymes such as creeatine kinase and the MB isoform (i.e., CK and CK/MB) which do not reach serum levels until several hours post-infarction (approx. 19 hours), myoglobin levels can be expected to peak within 6 to 9 hours. The Myoglobin Enzyme Immunoassay provides a rapid, sensitive, and reliable assay for the quantitative measurement of myoglobin in serum. The antibodies developed for the test will determine a minimal concentration of 5.0 ng/ml, and there is no cross-reactivity with related cardiac or skeletal enzymes. PRINCIPLE OF THE TEST The Myoglobin ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a unique monoclonal antibody directed against a distinct antigenic determinant on the myoglobin molecule. Mouse monoclonal anti-myoglobin antibody is used for solid phase immobilization (on the microtiter wells). A goat anti-myoglobin antibody is in the antibody-enzyme (horseradish peroxidase) 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 a 45 minute incubation at room temperature, the wells are washed with water 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 directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at

 Open the catalog to page 1
*Prices are pre-tax. They exclude delivery charges and customs duties and do not include additional charges for installation or activation options. Prices are indicative only and may vary by country, with changes to the cost of raw materials and exchange rates.