Human MIP4alpha(Macrophage Inflammatory Protein 4 Alpha) ELISA Kit

Mfr.No.

E-EL-H0031

Antigen

MIP4α

Alternative Names

CCL26 ; SCYA26 ; Eotaxin-3 ; IMAC ; MIP-4a ; TSC-1

Background

This kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human AGC .Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human AGC 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 Human AGC ,biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The OD value is measured spectrophotometrically at a wavelength of 450nm. The OD value is positively associated with the concentration of Human AGC . The concentration of Human AGC in the samples can be calculated by comparing the OD of the samples to the standard curve.

Description

As the OD 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)	O.D	Average	Corrected
2000	2.428 2.436	2.432	2.369
1000	1.715 1.721	1.718	1.655
500	0.9379999999999999 0.902	0.9199999999999999	0.857
250	0.465 0.479	0.472	0.409
125	0.255 0.253	0.254	0.191
62.5	0.176 0.154	0.165	0.102
31.25	0.115 0.115	0.115	0.052
0	0.056 0.07000000000000001	0.063	--

Specificity

This kit recognizes Human MIP4α in samples. No significant cross-reactivity or interference between Human MIP4α 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 ELISA 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	1 vial, 10 mL	4℃(shading light)
Stop Solution	1 vial, 10 mL	4℃
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Detection Method

Colormetric

Detection Range

31.25--2000pg/mL

Linearity

Samples were spiked with high concentrations of Human MIP4α 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 (%)	95-107	98-112	99-112
1:2	Average (%)	101	105	106
1:4	Range (%)	89-101	84-97	86-100
1:4	Average (%)	96	89	93
1:8	Range (%)	86-97	81-90	85-97
1:8	Average (%)	92	85	92
1:16	Range (%)	87-102	85-97	83-98
1:16	Average (%)	93	90	89

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MIP4α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MIP4α 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)	92.65	190.18	797.91	91.39	201.23	739.52
Standard	deviation	5.06	9.30	39.10	5.47	9.28	34.39
C V (%)	5.46	4.89	4.90	5.99	4.61	4.65

Recovery

The recovery of Human MIP4α 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)	90-102	96
EDTA plasma (n=5)	90-103	95
Cell culture media (n=5)	86-99	91

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

18.75 pg/mL

Uniprot	P55774

Manual

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Human MIP4alpha(Macrophage Inflammatory Protein 4 Alpha) ELISA Kit

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