Cat No.eia06148h        

Human Orexin B(OXB) ELISA Kit

Dear customers, thank you for choosing our products. This product is suitable for in vitro qualitative detection of human serum, plasma or cell culture supernatant and organizations in the natural and recombinant OXB 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 specimens.

4. Add horseradish peroxidase to label the avidins 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 specimens.

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

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 peroxidases combining with antibodies which shows higher sensitivity and amplification effect comparing with traditional direct HRP-Antibodies.

【Detection principle of Human Orexin B(OXB) ELISA kit】

This experiment use double-sandwich elisa technique and the ELISA Kit provided is typical. The pre-coated antibody is human OXB 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 Avidin-peroxidase  

【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.

【Necessary for testing their own testing supplies and reagents】

1. Centrifuge tube (capacity of 1.5ml, 5ml, and so on).

2. Disposable tip (range of 0.5-10μl-20μl, 20-200μl, 200-1000μl).

3. Pure water or distilled water.

4. Coordinate paper.

5. Absorbent paper.

6. EDTA, sodium citrate, heparin

 【Measures for the samples】

1.   Serum: Put the collected whole blood in refrigerator at 4℃ for the night. Then centrifuge it for 10min at 1000-3000rpm. Take supernatant tested immediately or put samples at -20℃( for 1-3 months) or -80℃ (for 1-3 months) for storage.

2.   Plasma: Take EDTA, sodium citrate and heparin as anticoagulant. Add the plasma and mix them well. Centrifuge mixture for 10min at 1000-3000rpm. Take supernatant tested immediately or put samples at -20℃( for 1-3 months) or -80℃ (for 1-3 months) for storage.【Note】 

1.  The kit should be kept at 2-8℃ before being used. Except the redissolved standard samples, other Ingredients must not be frozen.

2. For the concentrated biotinylated Human OXB antibody, the concentrated enzyme-conjugates have small size. Bumping or potential inversion of the tubes during transportation may cause the liquid sticking to wall or cap. Thus, the tubes should be shaken manually or centrifuged for 1 min at 1000rpm to shake off the adherent liquid down to the tube bottom.

3.  Concentrated washing buffer may crystallize a little. Use water bath to help the dissolution during diluting process. The crystals must be totally dissolved when preparing washing buffer.

4. During testing process, the human OXB lyophilized standard sample shall be single-use and must not be divided. The sample will quickly inactivate after being dissolved because of its lower concentration.

_{以上仅供参考，因实验室系统升级，各检测指标说明书有所更新，最终操作说明请
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