Human C5a(Complement Component 5a) CLIA Kit

Mfr.No.

E-CL-H0174

Antigen

C5a

Alternative Names

C5a

Background

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human ANG1 Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ANG1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain

Description

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration(pg/mL)	RLU 1	RLU 2	Average RLU	Corrected RLU
1000	53570	53780	53675	53650
500	20720	24258	22489	22464
250	10461	9961	10211	10186
125	4899	4903	4901	4876
62.5	2609	2297	2453	2428
31.25	1361	1201	1281	1256
15.63	691	725	708	683
0	24	26	25	--

Specificity

This kit recognizes Human C5a in samples. No significant cross-reactivity or interference between Human C5a and analogues was observed.

Assay Time

4.5h

Components

An unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro CLIA Plate(Dismountable)	8 wells ×12 strips	-20℃, 6 months
Reference Standard	2 vials
Concentrated Biotinylated Detection Ab (100×)	1 vial, 120 μL
Concentrated HRP Conjugate (100×)	1 vial, 120 μL	-20℃(shading light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	4℃, 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 A	1 vial, 5 mL	4℃ (shading light)
Substrate Reagent B	1 vial, 5 mL	4℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Detection Method

Chemiluminescence

Detection Range

15.625--1000 pg/mL

Linearity

Samples were spiked with high concentrations of Human C5a and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	98-114	100-118	98-112
1:2	Average (%)	105	108	104
1:4	Range (%)	100-112	101-113	84-95
1:4	Average (%)	106	107	90
1:8	Range (%)	93-107	103-117	95-108
1:8	Average (%)	99	109	101
1:16	Range (%)	88-104	98-115	87-100
1:16	Average (%)	95	105	92

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human C5a were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human C5a 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 (pg/mL)	52.90	130.31	464.60	48.44	133.53	510.37
Standard deviation	5.70	9.17	44.97	4.92	14.93	38.48
C V (%)	10.78	7.04	9.68	10.16	11.18	7.54

Recovery

The recovery of Human C5a 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=5)	96-114	104
EDTA plasma (n=5)	91-106	97
Cell culture media (n=5)	97-110	104

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

9.38 pg/mL

Manual

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SKU	Supplier No.	Size	Your price
BHE10300164	E-CL-H0174	96T	$682	Add to Cart
BHE10300164	E-CL-H0174	96T*5	$3069	Add to Cart
BHE10300164	E-CL-H0174	96T*10	$5797	Add to Cart
BHE10300164	E-CL-H0174	48T	$545.60	Add to Cart

Human C5a(Complement Component 5a) CLIA Kit

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