Albumin ELISA Kit - ALPCO Diagnostics

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Offered in the US by ALPCO www.alpco.com | Phone: (800) 592-5726 | Fax: (603) 898-68549. ASSAY PROCEDUREProcedural notes• Do not interchange different lot numbers of any kit component withinthe same assay.• Quality control guidelines should be observed.• Incubation time, incubation temperature and pipetting volumes of thecomponents are defined by the producer. Any variation of the testprocedure, which is not coordinated with the producer, may influencethe results of the test. Immundiagnostik AG can therefore not be heldresponsible for any damage resulting from wrong use.• The assay should always be performed according the enclosed manual.Test procedureWash the precoated PLATE (microtiter plate) 5 x with 250 µl diluted washbuffer. After the final washing step, the inverted PLATE should be firmlytapped on absorbent paper to remove excess solution. This is valid for allfollowing washing steps. Carry out each determination in duplicate forstandards, controls and samples.1. Add 100 µl STD (Standard), CTRL (Controls) and diluted patientsamples into respective wells.2. Incubate for 1 hour shaking on a horizontal mixer at roomtemperature.3. Decant the content of the PLATE and wash the wells 5 x with 250µldiluted wash buffer.4. Add 100 µl diluted conjugate in each well.5. Incubate for 1 hour shaking on a horizontal mixer at roomtemperature.6. Decant the content of the PLATE and wash the wells 5 x with 250µldiluted wash buffer.7. Add 100 µl SUB (TMB substrate) in each well.8. Incubate for 10-20 minutes at room temperature.9. Add 50 µl STOP (stop solution) in each well and mix shortly.10. Determine absorption immediately with an ELISA reader at 450 nmagainst 620 nm (or 690 nm) as reference. If no reference wavelength isavailable, read only at 450 nm. If the extinction of the highest standardexceeds the measurement range of the photometer, absorption mustbe measured immediately at 405 nm against 620 nm (or 690 nm) asreference.For reference use onlyFor research use only
Offered in the US by ALPCO www.alpco.com | Phone: (800) 592-5726 | Fax: (603) 898-685410. RESULTSWe recommend the 4-parameter-algorithm for evaluation of the test results.Alternatively, a point-to-point-calculation or a spline-algorithm can be used.Before the automatic evaluation of the results, the plausibility of the pairs ofvalues should be examined. If this option is not available with the usedprogram, a control of the paired values should be done manually.Typical calibration curveFor reference use onlyFor research use onlyConcentration [ng/ml] 800 200 50 12.5 0OD mean values 1.353 1.114 0.687 0.269 0.118The data is for demonstration only and cannot be used for the evaluation oftest results.
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Albumin ELISA Kit - ALPCO Diagnostics

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