Human Compiement C5 ELISA Kit

Protocol Booklet

Key features and details

Specification: 96 Test
Sensitivity: 0.01 ng/ml (50 μl)；0.18 ng/ml (10 μl)
Standard Curve Range: 1.37~1000 pg/ml
Standard Curve Gradient: 7 Points/3 Folds
Number of Incubations: 2
Sample Volume: 50 μl/10 μl
Assay type: Sandwich Elisa
Operation Duration: 120min

Brand:

CAT.NO. : AEHY0007

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Size:

96T

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Product Details

Background

Human complement component C5a (C5a) is an enzymatically generated glycoprotein that belongs to a family of structurally and functionally related proteins known as anaphylatoxins. C5a is a 74 amino acid (aa) peptide that is created by the activity of C5a convertase on the C5 alpha -chain. Human C5a has four alpha -helices plus three intrachain disulfide bonds that create a triple loop structure. In serum, proteolytic processing removes the C-terminal arginine, creating a low activity C5a desArg74 molecule. Human C5a is 60% and 54% aa identical to mouse and rat C5a, respectively. C5a binds to a signaling G-protein coupled receptor (C5aR/CD88) and a non-signaling GPCR termed C5L2. Activation of Cd88 results in neutrophil chemotaxis and endothelial cell activation. It also triggers an oxidative burst in macrophages and neutrophils, and induces release of histamine in basophils and mast cells.

typical data

ng/ml	O.D.		Average	Corrected
0.00	0.0061	0.0057	0.0059
1.37	0.0318	0.0300	0.0309	0.0250
4.12	0.0813	0.0794	0.0804	0.0745
12.35	0.2275	0.2267	0.2271	0.2212
37.04	0.6468	0.6343	0.6406	0.6347
111.11	1.5860	1.5560	1.5710	1.5651
333.33	3.2300	3.0340	3.1320	3.1261
1000.00	4.1306	3.9187	4.0247	4.0188

Precision

	Intra-assay Precision			Inter-assay Precision
Sample Number	S1	S2	S3	S1	S2	S3
Sample Number	22	22	22	6	6	6
Average（ng/ml）	17.8	92.6	287.7	16.6	85.8	261.2
Standard deviation	0.7	4.5	15.2	0.7	4.8	16.9
Coefficient of variation（%）	3.7	4.9	5.3	4.5	5.6	6.5

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of human Complement C5 into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 91% to 118% with an overall mean recovery of 101%.

Sample Values

Sample Matrix

Sample

Evaluated

Range

(μg/ml)

Detectable

Mean of Detectable

(μg/ml)

Serum

1.79-58.93

100

25.41

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of Complement C5 in this assay. No medical histories were available for the donors.

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