ALerCHEK's
Canine/ Feline
Allergen
Specific IgE Assay
Intended Use:
To qualify and quantify allergen specific Immunoglobulin E (IgE)
Principle of Procedure:
Solid phase capture sandwich ELISAassay
using an allergen coated
microwell
format.
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.
Specimen Dilution:
Dilute
each serum or plasma specimen to be tested 1:5 with the specimen
diluent provided.
Materials Supplied:
Two allergen coated microwell
strip plates (12 x 8) with plastic frame
Two
60mL vials of specimen diluent green
Two
60mL vials of 15X wash buffer concentrate
Two 12mL
vials of rabbit anti-canine/feline IgE (blue solution)
Two 12mL vials of
HRP conjugated goat anti-rabbit IgG (red solution)
One
30
mL vial of TMB/ peroxide color developer
Two 12 mL vials of sulfuric acid termination reagent
(0.5N)
Limitations of the
Procedure:
No single assay
should be used as the only basis for arriving at a diagnostic conclusion.
Reproducibility:
C.V. 6%-10% depending upon the region of
the standard curve.
Assay Procedure:
*Allow each reagent to
reach room temperature before use, dilute specimen and incubate at 4°C for 18 - 24
hours before use
1. Arrange the microwell strips in plastic
frame according to schematic provided
2. Add 100uL of diluted specimen to each
microwell
3. Incubate at room temperature for 2 hours
4. Decant and wash each microwell five times
with wash buffer (dilute buffer 1:15 with reagent grade water)
5. Add 100uL of
R-3 (blue solution) and then 100 uL of R-4 (red solution)
to each well
6. Incubate at room temperature for 3 hours
7. Decant and wash as in step 4
8. Add 100uL of TMB/peroxide substrate and
incubate at room temperature for 30 minutes
9. Terminate the reaction with 100uL of 1.0N
sulfuric acid
10. Zero plate reader on the negative control
well
11. Read with a standard microwell plate
reader at 450 nm
12. Interpret results with the following
scoring scheme:
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 mite |
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 Hair |
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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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