Cat No.eia05116m       

Mouse α-L-iduronidase(IDUA) ELISA Kit

Dear customers, thank you for choosing our products. This product is suitable for in vitro qualitative detection of Mouse serum, plasma or cell culture supernatant and organizations in the natural and recombinant IDUA concentration. Detection of other special sample please contact our technical support. The kit is for research use only. Please read the instructions carefully before using and check the kit components. If you have any questions, please contact SANCHEZ INC You will get our full range of services.

This kit employs  Double Antibody Sandwich Technique. The principle of Double Antibody Sandwich is based on characteristics of the tested antigen with more than two valances which can identify coated antibody  and detection antibody at same time. The specific process is as follows:

1. Connect the specific antibodies and solid phase carriers to form immobilized antibodies. Wash out uncombined antibodies and impurities. Seal the rest binding sites with irrelevant proteins.

2. Join under test with immobilized antibodies for contact reaction. After a while, combine antigens in and antibodies on carriers into the antigens complex. Wash out uncombined antibodies and impurities.

3. Add biotin  labeling  antibodies to combine with the antigens on immune complexes. Wash out the uncombined biotin  labeling antibodies thoroughly. The enzyme amount  on the carrier is now positively related to the amount of the tested substance  in specimen s.

4. Add horseradish peroxidase  to label the avidin s and incorporate them with the biotin  labeling antibodies. Wash out the incorporated enzyme markers  thoroughly. The enzyme amount  on the carrier is now positively related to the amount of the tested substance  in  specimen s.

5. Add substrates for coloring, and compute the concentration of specimen s.

Note: an antibody molecule can be marked on several biotin molecules and a biotin molecule can be connected with a  HRP-Avidin to form numbers of horseradish peroxidase s combining with antibodies which shows higher sensitivity and amplification effect comparing with traditional direct HRP-Antibodies.

【Detection principle of Mouse α-L-iduronidase(IDUA) ELISA kit】

This experiment use double-sandwich elisa  technique  and the ELISA Kit provided is typical. The pre-coated antibody is Mouse IDUA monoclonal antibody and the detecting antibody is polyclonal antibody with biotin labeled. Samples and biotin labeling antibody are added into ELISA plate wells and washed out with PBS or TBS. Then A vidin-peroxidase conjugates are added to ELISA wells in order; Use TMB substrate for coloring after reactant  thoroughly washed out by PBS or TBS. TMB turns into blue in peroxidase catalytic and finally turns into yellow under the action of acid. The color depth and the testing factors in samples are positively correlated.

【Kit composition】

name                    96 Tests  48 TestsStorage

1．antibody precoated plate    8×12      8×64/-20℃

2．Mouse IDUA Standards   2 vial      1 vial4/-20℃

3．Biotinylated antibody(1:100)    1 vial     1 vial4/-20℃

4．Enzyme conjugate(1:100) 1vial      1 vial4/-20℃

5．Enzyme diluent                1vial  1vial        4/-20℃

6．antibody diluent               1vial  1 vial4/-20℃

7．Standard diluent               1vial 1 vial      4/-20℃

8．Sample diluent                1vial  1 vial   4/-20℃

9．Washing buffer (1:25)         1 vial 1 vial    4/-20℃

10．Colour Reagent A  1 vial 1 vial   4/-20℃

11．Colour Reagent B1 vial 1 vial   4/-20℃

12．Colour Reagent C  1 vial 1 vial   4/-20℃

13．Instruction    1 set 1 set   RT

Note：

RT：Room temperature

Standard：Frozen dried

Colour Reagent A: Avoid light

【Necessary for testing their own test facilities and equipment】

1. Microplate reader (450nm detection wavelength filter, 570nm or 630nm correction wavelength filters).

2. Washer (adjustable amount of liquid injection to ensure that each well 350μl lotion without overflow).

3. Clean benches, biological safety cabinets, fume hoods.

4. High-precision single-channel dispenser (range 0.5-10μl-20μl, 20-200μl, 200-1000μl).

5. High-precision multi-channel plus liquid (8 or 12, the range of 50-300μl of).

6. 37℃ incubator.

7. Low temperature centrifuge.

8. Refrigerators (4℃, -20℃, -86℃).

9. Analytical balance.

10. Scissors, tweezers, pliers, and so on.

11. Swirl mixing device, low-frequency oscillator, and so on.