Detection kit of hexamethylene tetramine, detection method and application
A technology of hexamethylenetetramine and detection kits, applied in the direction of measuring devices, instruments, scientific instruments, etc.
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Embodiment 1
[0045] Example 1 Preparation of Hexamethylenetetramine Complete Antigen
[0046] Prepare 5ml of complex phosphate buffer solution with a pH value of 7.4, an ion concentration of 100 mmol, and 200 mg of bovine serum albumin (carrier protein); weigh 100 mg of hexamethylenetetramine and dissolve it in 400 μl of methanol. Drop into the prepared compound phosphate buffer; then add 3ml of 25% glutaraldehyde aqueous solution by volume, mix well, let stand at room temperature for 1 hour, refrigerate overnight at 4°C, and dialyze continuously with phosphate buffer for 5 day, the medium was changed once a day to obtain the complete antigen of hexamethylenetetramine (complete antigen HMTA-BSA).
Embodiment 2
[0047] Example 2 Identification of Hexamethylenetetramine Complete Antigen
[0048] Precisely prepare standard solutions of hexamethylenetetramine and bovine serum albumin (BSA) with deionized water. Prepare the conjugate HMTA-BSA standard solution with deionized water, measure the protein concentration by Kjeldahl method, and make the protein concentration of the conjugate solution consistent with the BSA protein concentration. Scan the spectrum to judge whether the coupling is successful. See the result figure 1 and figure 2 .
[0049] The results show that: the absorbance value corresponding to the maximum absorption wavelength of the HMT antigen is significantly different from that of HST and BSA, which is determined by figure 1 and figure 2 The comparison shows that after hexamethylenetetramine is coupled with the carrier protein, the ultraviolet spectrum scanning pattern of the conjugate is compared with that of the carrier protein BSA, and the spectral curve ch...
Embodiment 3
[0050] The preparation of embodiment 3 hexamethylenetetramine coated antigen
[0051] Accurately weigh 5 mg of hexamethylenetetramine and dissolve in 300 μl of methanol, then accurately weigh 12 mg of ovalbumin (OVA) and dissolve in 2.5 ml of phosphate buffered saline (PBS) with a concentration of 0.1 mol / L and a pH value of 7.4 in; slowly drop the methanol solution with hexamethylenetetramine into the ovalbumin phosphate buffer solution, then add 1.5ml of 25% glutaraldehyde aqueous solution by volume, mix well, and let stand at room temperature for 1 Hours, 4 ° C refrigerator overnight; put it into a dialysis bag, 4 ° C in 0.01mol / L, pH value of 7.4 phosphate buffer solution for 5 days, change the medium once a day; the obtained hexamethylene Tetramine-coated antigen (coated antigen HMTA-OVA), the antigen was divided into 1.5ml centrifuge tubes with carbonate buffer (concentration: 0.05mol / L, pH: 9.6) at a concentration of 0.75mg / ml stored at -80°C for later use.
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