[DLDevelop] Bovine Complement Component 3 (C3) ELISA Kit

[DLDevelop] Bovine Complement Component 3 (C3) ELISA Kit

Product name:	Bovine Complement Component 3 (C3) ELISA Kit
Method:	Sandwich
Synonyms:	ASP; CPAMD1; Complement Protein C3; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1; C3a anaphylatoxin; Acylation stimulating protein
Detection range:	7.81-500ng/mL
Reactivity:	C3
Size:	96T/48T
Quality guarantee period:	for 12 months
Catalog number:	DL-C3-b (traditional)	DLR-C3-b (ready-to-use)
Assay length	1-4.5Hours	1-3.5Hours
Advantages:	Competitive price.High sensitivity.High stability.12 months shelf life.	Pre-diluted Detection Reagent A and BReduction in the number of steps when conducting the testAll the reagents can be stored at 4℃Faster reaction compare to other brands12 months shelf life

Item	Standard	Test
Description	The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of C3 in bovine serum, plasma, tissue homogenates or other biological fluids.	Conform
Identification	Colorimetric	Positive
Composition	Traditional ELISA Kit	Ready-to-Use ELISA KIT	Conform
Pre-coated, ready to use 96-well strip plate 1	Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2	Plate sealer for 96 wells 2
Standard 2	Standard 2
Diluents buffer 1×45mL	Standard Diluent 1×20mL
Detection Reagent A 1×120μL	Detection Solution A 1×12mL
Detection Reagent B 1×120μL	Detection Solution B 1×12mL
TMB Substrate 1×9mL	TMB Substrate 1×9mL
Stop Solution 1×6mL	Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL	Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1	Instruction manual 1

Test principle

The microtiter plate provided in this kit has been pre-coated with an antibody specific to C3. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to C3. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain C3, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of C3 in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Recovery

Matrices listed below were spiked with certain level of recombinant C3 and the recovery rates were calculated by comparing the measured value to the expected amount of C3 in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	80-102	91
EDTA plasma(n=5)	81-98	89
heparin plasma(n=5)	80-89	84

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of C3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1：4	1：8	1：16
serum(n=5)	82-96%	83-98%	81-99%	93-101%
EDTA plasma(n=5)	88-101%	86-95%	90-102%	80-93%
heparin plasma(n=5)	80-91%	82-90%	95-104%	79-95%

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level C3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level C3 were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Stability

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage conditions.
Note:
To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37℃;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37℃;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
8. Add 50µL Stop Solution. Read at 450nm immediately.