Human
Allergen Specific IgA Assay
Intended Use:
To quantitate human Immunoglobulin A (IgA) specific for common allergens.
Principle of
Procedure:
Solid phase
capture sandwich ELISA assay using allergen
coated microwells (over 400 types available).
Shelf Life:
The expiration date for the package and each component is stated on the
label(s). Store all components at 2º- 8° degrees C. Do not freeze all or in part.
Patient and Standard Dilutions:
Dilute each serum or plasma specimen to be tested 1:10 with the Specimen Diluent provided. Use Specimen Diluent alone as a zero control or blank.
Materials Supplied:
Allergen coated microwell strips
8X12 with
plastic frame
HRP conjugated goat anti-human IgA - 12mL
TMB/peroxide substrate color developer - 12mL
Specimen Diluent - 2X 60mL
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:
0.00 - 5.6 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 buffer 1:15 with
reagent grade water)
4. Add
100uL of HRP conjugated goat anti-human IgA 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
O.D. @450nm: Class:
0.000-0.250 Class 0
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