Polyamines are generally believed to function both in protein synthesis and DNA synthesis leading to control cell proliferation. In 1971, Russel firstly reported that total amount of urinary polyamines elevated in cancer patients. And quantitative kit of urinary polyamines were already developed and utilized as a general biochemical examination.
Recently two diacetyl-derivatives, N1,N12-diacetylspermine and N1,N8-diacetylspermidine, were found to be excreted in urine and form 0.6% and 1.4% of total polyamines respectively.
Comparing urine of diseased person with urine of healthy person, some reports suggested the possibility that diacetyl-derivatives correlate to the status of disease more closely than total amount of polyamines.
Our kit is convenient to quantify amount of urinary diacetylspermine by using ELISA method. This kit is only for research use, not for diagnosis.
Highly sensitive and specific
Strip type well, antigen pre-coated microplate
Assay range: 6.25~200nM
Calculation method of clinical data
In general , the amount of urine change easily by amount of water take or environment even in renal disease patients and healthy persons, and concentrations of excrement depend on their amount. The amount of urinary creatinine depends on the amount of muscle and their measurements correlate positively. Therefore, correct actual measurement of Diacetylspermidine concentration (nM) in urine with creatinine concentration (mg/dl).
Domain of standard curve : 9.375~600nM
Minimum measurement range for detection : 18.75nM
Minimun dilution number of urine sample : ?2
Minimum sensitivity for detection : 37.5nM
Within-run (n=20, 2 concentration) : CV(%) = 5.76, 2.58
Between-run (n=20, 2 concentration) : CV(%) = 6.76, 4.25
Recovery test : In the recovery study, recoveries between 96% and 108% were obtained for 2, 4, 8times dilutions of the sample urine
No influence to Hemoglobin 4500mg/dL Bilirubin 180mg/dL Glucose 1000mg/dL Ascorbic acid 40mg/dL
The Reagents should be stored at recommended temperature, -30Υ.
Do not use the reagents which is expired the date of usage.
Urine sample should be diluted more than 4 times with Dilution solution.
Do not leave the standard and Antibody for long time under room temperature.
The glassware for making coloring solution should be clean.
Since OPD (o-phenylendiamine) is harmful, handle with care.
Since Stop solution,1N H2SO4, is strong acid, handle with care.
The kit is constructed with well-adjusted combination in each lot.
Replaced combination among different lots may cause unexpected results.
This kit is only for research use.
Do not use for medicinal or any other purposees.
When using the reagents, take care to avoid them from touching to skin, mucous membrane, clothes, and getting into eye.
If the reagents happen to get into eye or mouth, wash out them and consult a doctor if you need.
After using the kit, wash your hand very carefully.
If you find that the packages of the reagents are broken or something wrong, do not use them.
When you store the reagents, make sure to avoid them from vaporizing, falling down.
After using the reagents, the packages should be discarded under the established rule.
We do not guarantee the quality of the packages and accompaniments if not used according this direction. ?Storage? All
|(1)||Antigen coated microtiter plate,96 wells||1 plate|
|(2)||Diacetylspermine standard||250l × 2|
|(3)||Antibody diluent||20mL × 1|
|(4)||Anti Diacetylspermine antibody concentrate(×100)||80l × 1|
|(5)||HRP-anti Rabbit IgG Antibody concentrate(×80)||60l × 1|
|(6)||OPD (o-phenylendiamine) tablets||2 tab.|
|(7)||Substrate solution||30 mL × 1|
|(8)||Stop solution||15 mL × 1|
|(9)||Wash buffer concentrate (×20)||30 mL × 1|
|(10)||Dilution plate||1 plate|
Russell DH:Increased polyamine concentration in the urine of human cancer patients.
Nature New Biol 233:144-145,1971
Hiramatsu K, Sugimoto M, Kamei S, Hoshino M, Kinoshita K, Iwasaki K, and Kawakita M:Determination of amounts of polyamines excreted in urine;demomstration of N1,N8-diacetylspermidine and N1,N12- diacetylspermine as components commonly occurring in normal human urine. J. Biochem., 117:107-112,1995
Sugimoto M, Hiramatsu K, Kamei S, Kinoshita K, Hoshino M, Iwasaki K, and Kawakita M
Significance of urinary N1,N8-diacetylspermidine and N1,N12- diacetylspermine as indicators of neoplastic diseases. J. Cancer Res. Clin. Oncol.,121:317-319,1995
Hiramatsu K, Sugimoto M, Kamei S, Hoshino M, Kinoshita K, Iwasaki K, and Kawakita M:Diagnostic and prognostic usefulness of N1,N8- diacetylspermidine and N1,N12- diacetylspermine in urine as novel markers of malignancy. J. Cancer Res. Clin. Oncol., 123:539-545,1997
Urinary Diacetylspermidine ELISA Kit