An experience laboratory technician completed this Fibrinogen ELISA from Abcam, catalog number AB108841, lot GR245646-10. The goal of the experiment was to determine the cross-reactivity of this kit for EDTA-prepared plasma samples and serum samples for M. mulatta. Samples were stored prior to thaw at -80 degrees Celsius.
Overall, the kit, like other Abcam ELISA kits, was easy to follow and straight-forward. The laboratory technician had no problem understanding any parts of the protocol.
Our test was performed in 2 basic steps. The first was to find the appropriate testable dilution in plasma and serum, and the second was to test the linearity and spike recovery.
To this end our first experiment tested 8 dilutions of both serum and plasma ranging from 1:10 to 1:640 (8 1:2 serial dilutions). In these dilution ranges, the serum was well within the OD range, however no linearity was observed in any part of the dilution scheme. The EDTA plasma was beyond the range of detection (under-diluted).
Our second experiment focused on the EDTA prepared plasma. We diluted n=2 rhesus plasma samples 1:10,000, 1:30,000, 1:90,000, and 1:270,000 and tested in duplicate (as were the standards). The rest of the assay was performed to protocol’s specifications. For both animals tested, 1:90,000 and 1:270,000 gave similar results, suggesting this is the linear range of the assay.
Absorbance measurements of the diluted samples were comparable between duplicates in both tests (pipetting error appeared to be at a minimum). We also performed a spike recovery in both pure diluent and diluted plasma. ~100% of spiked human Fibrinogen was detected in the 90,000 and 270,000 diluted samples.
Overall, we recommend this kit for detection of M. mulatta fibrinogen in EDTA-prepared plasma.
Overall, the kit, like other Abcam ELISA kits, was easy to follow and straight-forward. The laboratory technician had no problem understanding any parts of the protocol.
Our test was performed in 2 basic steps. The first was to find the appropriate testable dilution in plasma and serum, and the second was to test the linearity and spike recovery.
To this end our first experiment tested 8 dilutions of both serum and plasma ranging from 1:10 to 1:640 (8 1:2 serial dilutions). In these dilution ranges, the serum was well within the OD range, however no linearity was observed in any part of the dilution scheme. The EDTA plasma was beyond the range of detection (under-diluted).
Our second experiment focused on the EDTA prepared plasma. We diluted n=2 rhesus plasma samples 1:10,000, 1:30,000, 1:90,000, and 1:270,000 and tested in duplicate (as were the standards). The rest of the assay was performed to protocol’s specifications. For both animals tested, 1:90,000 and 1:270,000 gave similar results, suggesting this is the linear range of the assay.
Absorbance measurements of the diluted samples were comparable between duplicates in both tests (pipetting error appeared to be at a minimum). We also performed a spike recovery in both pure diluent and diluted plasma. ~100% of spiked human Fibrinogen was detected in the 90,000 and 270,000 diluted samples.
Overall, we recommend this kit for detection of M. mulatta fibrinogen in EDTA-prepared plasma.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
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投稿 Jun 07 2016