Human Allergen Specific IgM Assay
Intended Use:
To quantitate
human allergen specific Immunoglobulin M (IgM)
Principle of
Procedure:
Solid phase
capture sandwich ELISA assay using an
allergen coated microwell.
Shelf Life:
The expiration date for the package and each component is stated on the label(s). Store all components at 2°- 8° degrees C. Do not freeze all or in part.
Patient and Standard Dilutions:
Dilute each serum or plasma specimen to be tested initially 1:10 using the Specimen Diluent provided. e.g. 10ul of specimen into 90 ul Specimen Diluent. Use the Specimen Diluent alone as the Zero or Blank control well.
Materials Supplied:
Allergen coated microwell strips 8X12 with
plastic frame
HRP conjugated goat anti-human IgM - 12mL
IgM standard (pre-diluted 1:10,000)
TMB/peroxide substrate color developer - 12mL
IgM specimen diluent - 2x60mL
Sulfuric acid termination reagent (0.5N) - 12mL
15 X Wash buffer concentrate – 60mL
Limitations of the Procedure:
No single assay should be used as the only basis for
arriving at a diagnostic conclusion.
Dynamic Range:
0.20 mg/mL to 2.56 mg/mL.
Reproducibility:
C.V. 6%-10% depending upon the region of the
standard curve.
Assay Procedure:
*Allow each reagent to reach room temperature before use
1. Add
100uL of diluted specimen to each microwell
2. Incubate
at room temperature for 60 minutes
3. Decant
and wash each microwell four times with wash buffer (dilute wash buffer 1:15 with reagent grade water
prior to use)
4. Add
100uL of HRP conjugated goat anti-human IgM to each well
5. Incubate
at room temperature for 60 minutes
6. Decant
and wash as in step 3
7. Add
100uL of TMB/peroxide substrate and incubate at room temperature for 30 minutes
8. Terminate
the reaction with 100uL of 0.5N sulfuric acid
9. Zero
the microwell reader at 450nm using the Specimen Diluent zero control well
10. Determine the optical density (O.D.) of the remaining wells
Interpretation of Results:
O.D. at 450nm Class:
0.000-0.250 Class 0 (Negative)
0.250-0.350 Class I (Equivocal)
0.350-0.450 Class II (True Positive)
0.450-0.550 Class III
>0.550 Class IV
Allergens Available: (other allergens available upon request)
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INHALANT ALLERGENS: |
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A. alternata |
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Eastern White Pine |
Oat Grass |
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Yellow Dock |
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A. vulgaris (Mugwort) |
European Filbert |
Orchard Grass |
Yellow Pine |
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Abisdia ramosa |
English Plantain |
Ox-Eye Daisy |
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Acacia |
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English Walnut |
P. communis |
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Ambrosia psilostarch |
Epicoccum |
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P. pululans |
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American Beech |
F. oxysporum |
Paristaria |
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American Elm |
Fescue |
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Penicillium chrysogenum |
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American Hazelnut |
Fire Ant |
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Penicillium notatum |
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Aspergillus niger |
Fleas |
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Phoma glomerata |
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Aspergillus repens |
Fuarium |
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Pigeon Droppings |
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Aspergillus terrens |
G. candidum |
Pigsweed |
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Aureobasidium pullularia |
German Cockroach |
Poplar |
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Bahai Grass |
Giant Ragweed |
Pyrethrum |
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Baker's Yeast |
Giant Wild Rye |
Queen Palm |
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Bent Grass |
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Goat Hair |
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Rabbit Epithelial |
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Bermuda Grass |
Goose Feathers |
Ragweed |
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Birch |
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Guinea Pig Hair |
Rat Epithelial |
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Black Locust |
H. velgare (Barley) |
Red Mulberry |
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Black Oak |
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Hamster Epithelial |
Red Oak |
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Black Willow |
Helminthosporuium satiuum |
Redtop |
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Blue Gum |
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Hickory |
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Reed's Canary grass |
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Botrytis |
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Honey Myrtle |
Rhizopus nigricans |
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Box Elder |
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Horse Dander |
Roughmarsh Elder |
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Brewers Yeast |
House Dust |
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Russian Thistle |
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Brome |
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Human Hair |
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Rye Grass (perennial) |
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Brown Cockroach |
Italian Cypress |
S. aureus |
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C. Goldenrod |
Johnson Grass |
Sage Brush |
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C. hormodendrum |
June Grass |
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Sheep Sorrel |
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C. lunata |
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Kapoc |
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Sheeps Wool |
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Candida albicans |
Kochia |
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Short Ragweed |
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Cat Hair |
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Lamb's Quarters |
Smooth Alder |
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Cattle Hair |
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M. canis (ringworm) |
Spadicoides |
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Chicken Feathers |
Malaleuca |
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Stemphylium b. |
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Cladosporium |
Meadow Foxtail |
Sugar Maple |
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Cocklebur |
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Mesquite |
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Sweet Vernal Grass |
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Common Reed |
Mimosa |
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Sycamore |
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Cotton |
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Mites (D. farinae) |
T. aestivum (wheat pollen) |
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Cottonwood |
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Mites (D. pteronyssinus) |
T. canus (gut parasite) |
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Cultivated Wheat |
Mixed Feathers |
T. pisiformis (tapeworm) |
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Curvularia |
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Mixed Ragweed |
T. vulpis (gut parasite) |
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Curvularia lunata |
Mouse Epithelial |
Timothy |
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Dust Mites |
Mt. Cedar |
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Trichoderma veride |
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Mucor racemosus |
Trichophyton mentagrophytes |
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Dandelion |
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Mucor mucedo |
Trichothecium |
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Dog Dander |
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mugwort |
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Velvet Grass |
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Dog hairr |
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Nettle |
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White Ash |
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Duck Feathers |
Nigrospora |
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Wormwood |
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FOOD ALLERGENS: |
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Alfalfa |
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Haddock |
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Tomato |
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Alfalfa Meal |
Honeydew Melon |
Tuna |
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Almond |
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Kiwi Fruit |
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Turkey |
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Apple |
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Lactalbumin |
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Vanilla Bean |
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Atlantic Blue Mussel |
Lamb |
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Venison (Odecoileus) |
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Bacon |
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Lettuce |
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Walnut |
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Barley |
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Liver |
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Wheat |
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Bean Sprouts |
Lobster |
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White Bean |
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Beef |
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Mango |
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White Potato |
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Beet Pulp |
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Millet |
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Black Pepper |
Mushroom |
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Brewers Yeast |
Navy Bean |
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Broccoli |
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Oat |
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Buckwheat |
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Olive |
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Cabbage |
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Onion |
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Canola Meal |
Orange Pekoe Tea |
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Carrot |
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Orange |
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Casein |
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Parsley |
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Celery |
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Pea |
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Cheddar Cheese |
Peaches |
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Chicken |
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Peanut |
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Chocolate |
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Pear |
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Clam |
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Pecan |
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Cocoa |
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Pinto Bean |
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Cod |
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Pork |
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Coffee |
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Poultry |
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Corn |
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Raw Sunflower |
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Corn Gluten Meal |
Rice |
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Cottonseed Meal |
Rye |
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Crab |
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Salmon |
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Cultivated Oats |
Seaweed |
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Dry Milk |
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Scallop |
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Egg White |
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Soybean |
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Egg Whole |
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Shrimp |
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Egg Yolk |
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Strawberry |
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Fish Meal |
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Suger Beets |
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Flaxseed Meal |
Sunflower Meal |
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Flounder |
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Sweet Potato |
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Garlic |
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