Human C3b(Complement Fragment 3b) ELISA Kit Detection Range 0.31-20 ng/mL Sensitivity 0.19 ng/mL Product Details Properties Assay type Sandwich-ELISA Format 96T Assay time 3.5h Detection range 0.31-20 ng/mL Sensitivity 0.19 ng/mL Sample type &Sample volume serum, plasma and other biological fluids; 100μL Specificity This kit recognizes Human C3b in samples. No significant cross-reactivity or interference between Human C3b and analogues was observed. Reproducibility Both intra-CV and inter-CV are < 10%. Application This ELISA kit applies to the in vitro quantitative determination of Human C3b concentrations in serum, plasma and other biological fluids. Test Principle This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human C3b. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human C3b and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human C3b, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human C3b. You can calculate the concentration of Human C3b in the samples by comparing the OD of the samples to the standard curve. Kit Components and Storage An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received. Item Specifications Storage Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips48T: 8 wells ×6 strips -20℃, 6 months Reference Standard 96T: 2 vials48T: 1 vial Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL48T: 1 vial, 60 μL Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL48T: 1 vial, 60 μL -20℃(Protect from light), 6 months Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months Biotinylated Detection Ab Diluent 1 vial, 14 mL HRP Conjugate Diluent 1 vial, 14 mL Concentrated Wash Buffer (25×) 1 vial, 30 mL Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light) Stop Solution 1 vial, 10 mL 2-8°C Plate Sealer 5 pieces Manual 1 copy Certificate of Analysis 1 copy Technical Data: Precision Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human C3b were tested 20 times on one plate, respectively.Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human C3b were tested on 3 different plates, 20 replicates in each plate. Intra-assay Precision Inter-assay Precision Sample 1 2 3 1 2 3 n 20 20 20 20 20 20 Mean (ng/mL) 1.09 2.76 7.45 0.98 2.60 6.89 Standard deviation 0.06 0.16 0.35 0.05 0.15 0.31 CV (%) 5.50 5.80 4.70 5.10 5.77 4.50 Recovery The recovery of Human C3b spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices. Sample Type Range (%) Average Recovery (%) Serum(n=8) 88-102 94 EDTA plasma (n=8) 94-107 99 Cell culture media (n=8) 91-106 97 Target Information Database Links SwissProt: P01024 Synonyms C3b Research Area Immunology Assay Procedures 1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C 2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C 3. Aspirate and wash the plate for 3 times 4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times 5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C 6. Add 50μL Stop Solution 7. Read the plate at 450nm immediately. Calculation of the results
