Latex Allergen 4-in-1 ELISA

$1,020.00

The Latex 4-in-1 ELISA Assay Kit provides a rapid and easy method for the quantitative determination of four principal allergenic proteins (Hev b 1, Hev b 3, Hev b 5 and Hev b 6.02) in Hevea natural rubber and other products. The kit includes reagents necessary to analyse 4 different allergens (Hev b 1, Hev b 3, Hev b 5, Hev b 6.02) in up to 5 samples using one kit. The Eagle Biosciences Latex 4-in-1 ELISA Assay Kit is for Research Use Only and not for use in diagnostic procedures.

SKU: LAB431-K01 Categories: , , Tags: , ,

Latex Allergen 4-in-1 ELISA

The Latex Allergen 4-in-1 ELISA is For Research Use Only

Size: 4×96 wells
Dynamic Range: Hev b1, Hev b3: 10-1000 μg/L; Hev b 5: 5 – 100 μg/L; Hev b 6.02: 5 – 200 μg/L
Incubation Time: 2 hours
Sample Type: Rubber products
Sample Size: 25 µL
Controls Included


Assay Principle

The Latex Allergen 4-in-1 Assay Kit is based on the enzyme immunometric assay technique. Microtiter wells are pre-coated with one of 4 Hev b specific monoclonal antibody. Calibrators, controls and samples are pipetted into the wells and Hev b allergen is bound by immobilized antibody. After incubation, unbound material is removed by washing the wells. In the second incubation, horseradish peroxidase (HRP) labeled Hev b specific monoclonal antibodies are added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of Hev b allergen bound in the initial step. The color development is stopped and the intensity of the color is measured.


Related Products

Latex Allergen ELISA Kit – FITkit Hev b 1
Latex Allergen ELISA Assay Kit – FITkit Hev b 3
Latex Allergen ELISA Assay – FITkit Hev b 5

Additional Information

Assay Procedure


Allow all reagents to reach room temperature (25±5 °C) before use (at least 30 minutes). Perform the assay procedure at 25±5 °C. To help you easily find the reagents for specific Hev b analyze, the reagents labels are colour-coded.

  1. Dilute the PBS Wash Concentrate to500 mL with distilled water to make washing solution.
  2. Dispense 100 μL of Hev b 1, Hev b 3,Hev b 5 and Hev b 6.02 Assay Buffer into appropriate wells.
  3. Pipette 25 μL of Hev b 1, Hev b 3,Hev b 5 and Hev b 6.02 calibrators A-F into appropriate wells in duplicate.
  4. Pipette 25 μL of Hev b 1, Hev b 3,Hev b 5 and Hev b 6.02 control into appropriate wells in duplicate.
  5. Pipette 25 μL of samples into appropriate wells in duplicate.Optional: Cut out the provided plate overview and fill in the corresponding sample cells. Sample cells have been numbered.
  6. Cover the microtiter plate and incubate for 60 minutes at room temperature on a microtiter plate shaker (300rpm).
  7. Rinse the microtiter plate wells 4 times with 300 μL of washing solution.
  8. Strike the wells sharply onto absorbent paper or paper towels to remove all residual washing solution droplets.
  9. Dispense 100 μL of Hev b 1, Hev b 3,Hev b 5 and Hev b 6.02 Enzyme Conjugate into the appropriate wells.
  10. Cover the microtiter plate. Incubate the microtiter plate for 30 minutes at room temperature on a microtiter plate shaker (300 rpm).
  11. Rinse the microtiter plate wells 4 times with 300 μL of washing solution.
  12. Strike the wells sharply onto absorbent paper or paper towels to remove all residual washing solution droplets.
  13. Add 100 μL of HRP Substrate Solution at fixed time points into each well.
  14. Cover the microtiter plate. Incubate the microtiter plate for 15 minutes at room temperature on a microtiter plate shaker (300 rpm).
  15. Stop the reaction by adding 100 μLof Stopping Solution into each well at the same fixed points in time as in Step 12. Shake the microtiter plate for1-2 minutes to mix the solutions.
  16. Measure the absorbance at 414 nm or405 nm using a microtiter plate reader,preferably immediately but no more than 60 minutes after stopping the reaction.

Quality Control


  • The blank value (Calibrator A) must be ≤0.1.
  • Calibrators, controls and sample parallels variability (CV%=coefficient of variation)must be ≤ 15%.
  • Controls (Hev b 1 Control, Hev b 3 Control,Hev b 5 Control, Hev b 6.02 Control) should give results within the specified ranges given in a separate certificate of analysis enclosed in the kit.
  • Note – you have made four different Analyses –one for each Hev b.

Documents

Product Documents


Product Citations