ALerCHEK

is a developer, manufacturer, and distributor of immunoassay-based diagnostic test kits and components.

Apolipoprotein A1 (product insert):

Apo AI is primarily found in high density lipoprotein (HDL) particle. It serves the function of preventing the accumulation of cholesterol loaded macrophages which deposit on the arterial wall as foam cells. This is the prominent early feature of atherosclerotic lesion formation ultimately resulting in atherosclerosis. Apo AI, is a single polypeptide with a molecular weight of 28 Kd. Its primary function is to activate LCAT within the HDL complex, which catalyzes the esterification of cholesterol. This results in a more soluble cholesterol-HDL complex which increases the cholesterol transport capacity of the HDL particle for subsequent removal by the liver. Apo AI is therefore a convenient marker for assessing the cholesterol clearing capacity of the blood, and studies have clearly indicated that it is a better discriminator of angiographically documented coronary artery disease than HDL cholesterol.

Apolipoprotein B (product insert):

Over 90% of low density lipoprotein (LDL) particle is composed of Apo B. It serves the function of solubalizing cholesterol within the LDL complex, which in turn increases the transport capacity of LDL for subsequent deposit on the arterial wall. Apo B is therefore a convenient marker for assessing the cholesterol depositing capacity of the blood, and studies have clearly indicated it as a better discriminator of angiographically documented coronary artery disease than LDL cholesterol.

Lipoprotein(a) (product insert):

Lp(a), unlike Apo AI or Apo B, whose levels vary as a result of diet, exercise, etc. is predominantly a genetic trait whose level remains more or less constant after puberty. More than 13 phenotypes of Lp(a) have been identified having molecular weight of 300-800 Kd. It is bound to both HDL and LDL. Lp(a) interferes with plasminogen, the clot dissolving enzyme, which binds to the arterial endothelial lining. This in turn contributes to blood clot formation, and over a prolonged period of time would lead to significant damage to the coronary arteries. Levels of greater than 30mg/dl have been demonstrated to independently increase the risk of CHD by six fold.

C-Reactive Protein (product insert):

C-Reactive Protein (CRP) has been demonstrated to be a general indicator of major tissue damage. Hence, it can be used to indicate a stroke or heart attack because major blood vessels leading to the heart or brain are damaged and release large quantities of CRP during these disease states. CRP is a particularly useful indicator of CHD in women and in patients that demonstrate no other plasma circulating biochemical indicators.

Total Human IgE ELISA Assay

Intended Use:

To quantitate total human Immunoglobulin E (IgE)

Principle of Procedure:

Solid phase capture sandwich ELISA assay using a 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° C. Do not freeze all or in part.

Patient and Standards:

The Specimen diluent alone is used for a blank.

Materials Supplied:*

Anti-Human mouse monoclonal IgE coated microwell strips 8x12 with plastic frame –8 strips

HRP conjugated mouse monoclonal anti-human IgE -12mL

IgE standards (Low, Medium, High) –1 mL each

TMB/peroxide substrate color developer –12mL

IgE Specimen diluent -60mL

Sulfuric acid termination reagent (0.5N) –12mL

15 X Wash buffer concentrate – 60mL

* Quantities are representative of single kits - for double kits, multiply amounts by two.

Limitations of the Procedure:

No single assay should be used as the only basis for arriving at a diagnostic conclusion.

Dynamic Range:

0.0 IU/mL to 400 IU/mL.

Reproducibility:

C.V. 6%-10% depending upon the region of the standard curve.

Typical Standard Curve:

Assay Procedure:

Allow each reagent to reach room temperature before use

1. Add 10uL of specimen or 10ul of standard to each microwell. Use 100uL of specimen diluent alone for negative or blank well.

2. Add 90ul of Specimen diluent to each specimen or standard well.

3. Incubate at room temperature for 2 hours.

4. Decant and wash each microwell four times with wash buffer (diluted buffer 1:15 with reagent grade water). Firmly grip and pound the microwell strips upside down on dry folded paper towels to remove residual wash buffer after the last decanted wash.

5. Add 100uL of HRP conjugated mouse monoclonal anti-human IgE to each well

6. Incubate at room temperature for 2 hours.

7. Decant and wash as in "Step 3"

8. Add 100uL of TMB/peroxide substrate and incubate at room temperature for 30 minutes

9. Terminate the reaction with 100uL of 0.5N sulfuric acid

10. Zero the microwell reader at 450nm using the specimen diluent, zero control well

11. Determine the optical density (O.D.@450nm) of the remaining wells

12. Construct a standard curve using the O.D. values obtained for each of the standards

13. Interpolate the unknowns from the standard curve

Intended Use:

To qualify and quantify allergen specific Immunoglobulin E (IgE) Principle of Procedure:

Solid phase capture sandwich ELISA assay using a 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 specimen to be tested 1:5 with the specimen diluent provided.

Materials Supplied:

Two allergen coated microwell strips 12x8 with plastic frame

R-2 two 60mL vial of specimen diluent green
R-2 two 60mL vials of 15X wash buffer concentrate
R-3 one 60mL vials of rabbit anti-canine/feline IgE (blue solution)
R-4 one 60mL vials of HRP conjugated goat anti-rabbit IgG (red solution)

One 30 mL vials of TMB/ peroxide color developer

Two 12 mL vials of sulfuric acid termination reagent (1.0N)

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:

0 – 0.099 = NEGATIVE

0.100 – 0.199 = CLASS I

0.200 – 0.299 = CLASS II

0.300 – 0.399 = CLASS III