Description

Special Explanation

1. *If the kit is opened, Store the whole kit at 4°C. If the kit is not used up in 1 week. Store the Pre-Coated
Microplate, Standard ,Biotinylated Antibody and Streptavidin-HRP at -20°C, the rest reagents at 4°C,
please used up within 6 months.
*If the kit is not opened, store the whole kit: 4°C(short time storage, valid for 6 months); -20°C
(long-term storage, valid for 1 year). Avoid repeated freeze-thaw cycles.
2. Do not use the kit beyond the expiration date.
3. If the whole kit is stored at -20°C, place the kit at 4°C the day before the experiment.
4. After opening the package, please check that all components are complete.
5. The cap must be tightened to prevent evaporation and microbial contamination. The reagents
volume is slightly more than the volume marked on labels, please use accurate measuring
equipment and do not pour directly into the vial.
All kit components have been formulated and quality control tested to function successfully. Do not mix or
substitute reagents or materials from other kits, detection effect of the kit will not be guaranteed if utilized
separately or substituted.

Materials Required, Not Supplied

1. Microplate reader capable of measuring absorbance at 450 ± 10 nm.
2. High-speed centrifuge.
3. Electro-heating standing-temperature cultivator.
4. Absorbent paper.
5. Double distilled water or deionized water.
6. Single or multi-channel pipettes with high precision and disposable tips.
7. Precision pipettes to deliver 2 μL to 1 mL volumes.

Safety Notes

1. This kit is only used for lab research and development and should not be used for human or animals.
2. Reagents should be regarded as hazardous substances and should be handled carefully and correctly.
3. Gloves, lab coats, and goggles should always be worn to avoid skin and eyes coming into contact with
Stop Solution and TMB. In case of contact, wash thoroughly with water.

Test Principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in
this kit has been pre-coated with an antibody specific to Rat Smad3. Standards or samples are added to
the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat Smad3. Next,
Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After
TMB substrate solution is added, only those wells that contain Rat Smad3, biotin-conjugated antibody and
enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the
addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength
of 450nm ± 10nm. The concentration of Rat Smad3 in the samples is then determined by comparing the
OD of the samples to the standard curve.