Specimen Collection And Preparation; Assay Procedure; General Remarks - DRG EIA-2395 Instrucciones De Uso

5

SPECIMEN COLLECTION AND PREPARATION

Serum or plasma can be used in this assay.
Do not use haemolytic, icteric or lipaemic specimens.
Please note: Samples containing sodium azide should not be used in the assay.
5.1 Specimen Collection
Serum:
Collect blood by venipuncture (e.g. Sarstedt Monovette for serum), allow to clot, and separate serum by centrifugation at
room temperature. Do not centrifuge before complete clotting has occurred. Patients receiving anticoagulant therapy may
require increased clotting time.
Plasma:
Whole blood should be collected into centrifuge tubes containing anti-coagulant (e.g. Sarstedt Monovette with the
appropriate plasma preparation) and centrifuged immediately after collection.
5.2 Specimen Storage and Preparation
Specimens should be capped and may be stored for up to 24 hours at 2 °C - 8 °C prior to assaying.
Specimens held for a longer time should be frozen only once at -20 °C prior to assay. Thawed samples should be
inverted several times prior to testing.
5.3 Specimen Dilution
If in an initial assay, a specimen is found to contain more than the highest standard, the specimens can be diluted with
Standard 0 and re-assayed as described in Assay Procedure.
For the calculation of the concentrations this dilution factor has to be taken into account.
Example:
a) dilution 1:10: 10 µL sample + 90 µL Standard 0 (mix thoroughly)
b) dilution 1:100: 10 µL dilution a) 1:10 + 90 µL Standard 0 (mix thoroughly).
6

ASSAY PROCEDURE

6.1

General Remarks

− All reagents and specimens must be allowed to come to room temperature before use. All reagents must be mixed
without foaming.
− Once the test has been started, all steps should be completed without interruption.
− Use new disposal plastic pipette tips for each standard, control or sample in order to avoid cross contamination.
− Absorbance is a function of the incubation time and temperature. Before starting the assay, it is recommended that all
reagents are ready, caps removed, all needed wells secured in holder, etc. This will ensure equal elapsed time for
each pipetting step without interruption.
− As a general rule the enzymatic reaction is linearly proportional to time and temperature.
Version 10.0 2017/05 - vk
Leptin Sandwich ELISA EIA-2395
-
6 -