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 ELISA KIT

1.    What is the principle of Sincere Biotech TM ELISA Kit?

2.    What is the expiry date and store condition of  Sincere Biotech TM ELISA Kit?

3.    Are Positive or /and Negtive control  available for Colorimetric Sandwich ELISA ?

4.    Do you offer free sample or discounts on kits for testing?

5.    Why must I use polypropylene tubes for standard dilutions in certain assays?

6.    How are tissue samples prepared for assays if it is not described in the data sheet?

7.    What are the best suggestions to generate a standard curve with theSincere Biotech TM kits?

8.    Do Sincere Biotech TM kits kits with all body fluids, cell culture and tissue sample?

9.     What is the dilute principle of Sample ?

10.   How many samples could be detected with each ELISA Kit?

11. How to store the Samples?


1.    What is the principle of Sincere Biotech TM ELISA Kit?

Standard and samples are aspirated into the wells,Target protein is bound to monoclonal  capture antibody specific immobilized. The biotinylated secondary antibody specific is used for detection. Avidin-Biotin-Peroxidase Complex, ,TMB substrate solution is added, the color changes after adding acidic TMB Stop solution.Each step need sufficient wash.

2.    What is the expiry date and store condition of  Sincere Biotech TM ELISA Kit?

All the components in the kit should be stored up to 1 year at -20 and 3 months at 2-8,and should be kept according to the labels on vials. If the kit can not be used within 2weeks,please keep in -20℃.

3.    Are Positive or /and Negtive control  available for Colorimetric Sandwich ELISA ?

Sincere Biotech supply Positive or /and Negtive control for all Elisa kit item, Please inquire for specific ordering information.     

4.    Do you offer free sample or discounts on kits for testing?

In some occasional cases we may offer and 24T free sample and discounts.Please contact us directly .

 5.    Why must I use polypropylene tubes for standard dilutions in certain assays?

Certain proteins or analytes will bind to glass and polystyrene, but do not readily bind to the polypropylene tubes.

6.    How are tissue samples prepared for assays if it is not described in the data sheet?

1)    tissue (0.2g to 1g), rinsed in ice-cold physiological saline, the blood was removed, the filter paper drying, weighed, and placed in a small beaker of 5 or 10 ml

2)    The buffer(ph7.4, 0.01mol/LTris-HCL, 0.0001MOL/LEDTA-2Na, 0.01mol/L sucrose, 0.8% sodium chloride solution) or the buffer (0.86% cold saline) or PBS.

3)    Add the buffer,9 times volume to the tissue blocks, Shredded tissue blocks as soon as possible.

4)    Homogenized: with organization stamp mill in 10000 ~ 15000r/min, up and down grinding made the 10% homogenate.

5) put the 10% homogenate in low-temperature low-speed centrifuge ,3000r/min about centrifuged for 10 - 15 minutes, having the supernatant .

7.    What are the best suggestions to generate a standard curve with the Sincere Biotech TM kits?

The dilution scheme is detailed in the data sheet and should be followed accurately.

8.    Do Sincere Biotech TM kits kits with all body fluids, cell culture and tissue sample?

Unless specifically mentioned in themanal  for certain kinds of samples or interfering substances, most of the assay kits can be used with body fluids, plasma and serum samples, cell culture samples and tissue sample (snap frozen and fresh).

9.     What is the dilute principle of Sample ?

User needs to estimate the concentration of the target protein in the samples and select proper dilution factor so that the diluted target protein concentration falls near the middle of the linear regime in the standard curve. If samples are diluted before being added to the assay wells, please be sure to include the dilution factor for calculation of sample concentrations.

10.   How many samples could be detected with each ELISA Kit?

The kit contain sufficient materials to run ELISAs on 96 or 48 microplates. Specific vial volume of each component may vary.Please check carefully if all components is in the correct volume. If you have any other question, please see the product's Certificate of Analysis or contact us directly.

11.   How to store the Samples?

Samples to be used within 24hours may be stored at 4°C, otherwise samples must be stored at -20°C (≤3month) or -80°C(≤8 months) to avoid loss of bioactivity and contamination.



TECHNICAL HINTS AND LIMITATIONS

Ø Reagents are intended for research use only and are not for use in diagnostic or therapeutic procedures.

Ø Highly recommend that this product is handled only by those persons who have been trained in laboratory techniques and that it is used in accordance with the principles of good laboratory practice,due to all chemicals should be considered as potentially hazardous.

Ø Wear suitable protective clothing such as laboratory overalls, safety glasses and gloves. Care should be taken to avoid contact with skin or eyes. In the case of contact with skin or eyes wash immediately with water. See MSDS and/or safety statement(s) for specific advice.

Ø Handle all biological sample as potentially hazardous and capable of transmitting diseases.     

Ø Hemolyzed, hyperlipemic, heat-treated or contaminated samples may give  erroneous results.                       

Ø The waste disposal should be performed according to your laboratory  regulations.               

Ø Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.

Ø Do not pipette by mouth,and do not eat or smoke in areas where kit reagents or samples are handled.

      -Avoid contact of skin or mucous membranes with kit reagents or specimens.

      -Avoid contact of substrate solution with oxidizing agents and metal.

      -Avoid splashing or generation of aerosols.

Ø To avoid cross-contaminations, change pipette tips between reagent additions of each standard, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.

Ø Use polypropylene tubes for preparation of standard and samples. Do not use polystyrene tubes or sample plates.

Ø Do not reuse microwells or pour reagents back into their bottles once dispensed.

Ø Do not under any circumstances add sodium azide as preservative to any of the components.

Ø Caps and vials are not interchangeable. Caps should be replaced on the corresponding vials.

Ø Limited by the current condition and scientific technology, we can't completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.

Ø Kits from different manufacturers with the same item might produce different results, since we haven’t compared our products with other manufacturers.

Ø Please perform the experiment exactly according to the instruction attached in kit,Please use only the reagents supplied by manufacturer.

Ø Kits from different batches may be a little different in detection range, sensitivity and color developing time.

Ø There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.

Ø The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.

Ø Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.

Ø To ensure accurate results, proper adhesion of supplied covers during incubation steps is necessary.

Ø Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results.

Ø Incubation times, incubation temperature and pipetting volumes other than those specified may give erroneous results.

Ø Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.

Ø Since exact conditions may vary from assay to assay, a standard curve must be established for every run. If the standard is out of range, the results of the test samples are not reliable. The test should be repeated.

Ø The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

                                                                                                                   

 

 

 

 

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