Peptides immobilizing kit
Cat. No.: FNK-BCL-PIK-01
Size: 1kit
Storage: 4℃ (1 year)
Product outline
Peptide immobilizing kit is used for making peptide solid phase on normal ELISA plate. In general, peptides are difficult to immobilize on normal ELISA plate. The kit is consisted of reagents to make conjugate and coating buffer, and one can easily form peptide conjugate that can be immobilized on normal plate at high efficiency. The kit contains reagents enough to make conjugate three times of 250µg; of peptide. Five ELISA plates can be coated at 5 µg/mL of peptide at one time, total 15 plates.
Principle
Peptide forms conjugate with BSA, and immobilized on normal ELISA plate by using BSA as a foothold. It should be noted that antibody recognizes BSA is not used in this system. As coating buffer carbonate buffer is attached.
Product feature
1. Higher peptide solid phase than pepetide immobilizing amino ELISA plate*
2. Requires only 90 min for plate coating
3. Economical due to 3 times conjugate formation possible
4. Easy coating due to attached coating buffer
(*depends on the type and performance of amino plate)
How to use
1.Mixing of peptide and BSA solution
Peptide dissolved in PBS is mixed with BSA solution (1mg/mL. The basic mix ratio is 250µL of peptide （1mg/mL） and 250µL of BSA solution, but may need modification. As far as the condition of the total volume being 500µL and the BSA solution used being 250µL is kept constant, the peptide concentration can be varied within the range of 0.5 to 2mg/mL【note 1】.
2. Formation of conjugate
The following steps should be done at 4℃. Add 50µL of Reaction Solution to the mixed solution, and mix well by inverting the container, and let it stay for 30 sec. Repeat the same procedure 4 more times (totally 250µL of Reaction Solution is added). Then incubate for 60 min while mixing a few more times during the incubation.
3. Stopping reaction
Take out the container from refrigerator, and add all the powder from one tube of reaction Stop Agent into the container, and stop the reaction. Please keep the container opened during stop reaction since small gas 【note 2】 is generated by this reaction. Let it stay open until the gas generation stops.
4. Immobilization of peptide
Dilute 2 x Coating Buffer by the same volume of pure water. Dilute the conjugate solution to desired concentration by 1 x Coating Buffer, and immobilize peptide by adding 100µL/well of diluted conjugate (for 96-well plate). Incubation time is generally 30 min to 3 hrs.
5. Storage of conjugate
The prepared conjugate can be stored for 1 week at 4℃ 【note 3】.
【note 1】 The optimum mix ratio may be different according to peptide nature.
【note 2】 The gas is hydrogen. Although it is a small amount, please be careful of fire.
【note 3】 The storage time may be different according to peptide nature.
