Quantity | Sign | ingredient |
1×12×8 | MTP | The coated plate is detachable and the micropores are coated with anti-rabbit IgG (sheep, monoclonal). |
1×5ml | ANTISERUM | 5-HIAA antiserum Blue, ready to use, contains: antiserum (rabbit), phosphate buffer, stabilizer. |
1×5ml | BIOTIN | 5-HIAA Biotin Ready-to-use, blue, contains: phosphate buffer, stabilizer. |
1 x 0.2 mL | ENZCONJ CONC | Enzyme Concentrate (100×) Contains: Alkaline phosphatase-labeled avidin antibody (sheep), Tris buffer, stabilizer |
1×7×0.5ml | CAL AG | Standard AG 0; 0.4; 1.0; 2.75; 7.5; 20; 55 mg/L 0; 2.1; 5.25; 14.4; 39.4; 105; 288 μmol/L Ready to use, contains: 5-HIAA (methylation), stabilizer. |
1×2×0.5ml | CONTROL 1+2 LYO | Quality control 1+2, lyophilized powder, containing: human urine (normal and abnormal human urine), please refer to the reagent bottle label for specific concentration and allowable range. |
1×2ml | METHYLRAGE | Methylation reagent, yellow, ready to use, containing methylene chloride |
1 x 1 mL | HCL | HCl, ready to use, 0.1 M HCl |
1 x50ml | ASSAUBUF CONC | Detection buffer, concentrated (10 x) Contains: phosphate buffer, BSA, stabilizer |
1 x4ml | DILREAG | Diluent, ready to use, contains: N,N-dimethylformamide |
1 x50ml | WAHSBUF CONC | Washing liquid, concentrated (20 x) Contains: phosphate buffer, Tween, stabilizer |
1×9 | PNPP SUBS | PNPP substrate containing: p-nitrophenyl phosphate (PNPP) |
1×27ml | PNPP BUF | PNPP substrate buffer, ready to use, contains: diethanolamine, water, |
1×15ml | PNPP STOP | PNPP stop solution, ready to use, containing: 1M NaOH, 0.25M EDTA |
3× | FOIL | Sticky metal plate |
note | The 96-person test reagent can be divided into 3 independent experiments. The following volume is the amount of reagent required for one test with four coated plates (32 servings). If the customer wants to reduce the amount of the standard from 7 to 6, you can remove the standard G. The report range of the experimental results is correspondingly reduced to 3000 μg / L |
dilution | ingredient | Diluent | proportion | Remarks | Storage | stability | |
15ml | Detection buffer | 150ml | Double distilled water | 1:10 | Fully mixed | 2-8 ° C | 2 weeks |
Control product | 0.5ml | 0.1M HCl | Allow to stand for 15 min, avoid bubbles when mixing | ≤-20°C | 8 weeks | ||
15ml | detergent | 300ml | Double distilled water | 1:20 | Heat to 37 ° C to dissolve all crystals (if necessary) and dissolve thoroughly. | 2-8 ° C | 4 weeks |
60μL | Enzyme complex | 60ml | Detection buffer (diluted) | 1:101 | Temporary configuration, can only be used once, avoiding bubble generation during mixing | 18-25 ° C | 30min |
3 | PNPP substrate | 8ml | PNPP substrate buffer | Temporary configuration, can only be used once, free of bubbles when mixing | 18-25 ° C | 10min |
note | Do not methylate the standard, it has been methylated. |
1) | 20 μL of the control and the sample were separately added to the corresponding glass test tubes. |
2) | After completing the next steps, continue to operate in the fume hood. |
3) | 50 μL of the dilution was added to each tube. Mix on a vortex mixer. |
4) | 25 μL of methylation reagent was added to each tube. The test tube solution was mixed immediately after the addition of the reagent. Note: The yellow color of the reaction mixture should be stable. If the color disappears quickly, the amount of acid in the sample is too large. In this case, 20 μL of methylation reagent was added to the test tube. |
5) | Cover the tube and incubate for 20 min at room temperature (18-25 ° C) |
6) | Add 5 ml of diluted assay buffer to each tube. Cover the tube with a stopper and rotate it several times. Mix up to five times manually or with a mixer so that the liquid can mix well. |
7) | After completing this step, you can not operate under the fume hood. |
8) | 50 μL of the supernatant was aspirated and an ELISA experiment was performed immediately. The supernatant was stable for 1 h at room temperature. |
1) | 50 μL of the standard, methylation control and methylated patient samples were added to the corresponding microwells. |
2) | 50 μL of 5-HIAA biotin was added to each well. |
3) | 50 μL of 5-HIAA antiserum was added to each well. |
4) | Cover the plate and carefully shake the cover plate. Incubate overnight (16-20 h) at 2-8 °C. |
1) | The metal plate was removed, the reaction solution in the well was discarded, and the plate was washed 3 times with 250 μL of the washing solution, and patted dry on the absorbent paper. |
2) | Add 150 μL of temporarily configured enzyme conjugate to each well |
3) | Use a new sticky metal cover. Incubate for 120 min on an orbital shaker (500 rpm) at room temperature (18-25 ° C). |
4) | The enzyme conjugate was added to the microwells approximately 10 minutes before the incubation of the genus. |
5) | The metal plate was removed, the reaction solution in the well was discarded, and the plate was washed 3 times with 250 μL of the diluted washing solution, and patted dry on the absorbent paper. |
6) | When adding the substrate and stop solution, if necessary, an 8-channel pipette can be used for dosing. The substrate and stop solution must be applied at the same time interval to avoid bubble generation. |
7) | 200 μL of temporarily configured PNPP substrate solution was added to each well. |
8) | Incubate for 60 min on an orbital shaker at room temperature (18-25 ° C). |
9) | 50 μL of PNPP stop solution was added to each well to terminate the reaction. Gently shake the coated plate to mix the ingredients. |
10) | The OD value (reference wavelength: 600-650 nm) was measured at 405 nm within 60 min after the addition of the stop solution. |
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Mg/24h | Molmol/day | ||
5-HIAA | 6 to 10 | 31.5~52.5 | |
Streptococcus Thermophilus,Streptococcus Thermophilus Powder,Streptococcus Thermophilus Probiotic,Streptococcus Thermophilus Bacteria
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