Rat UCP-1(Uncoupling Protein 1, Mitochondrial) ELISA Kit

Mfr.No.

E-EL-R1146

Antigen

UCP-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 A-CoA .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 A-CoA 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 A-CoA ,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 A-CoA . The concentration of A-CoA 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(ng/mL)	O.D	Average	Corrected
10	2.351 2.379	2.365	2.3
5	1.49 1.496	1.493	1.428
2.5	0.803 0.765	0.784	0.719
1.25	0.438 0.442	0.44	0.375
0.63	0.245 0.235	0.24	0.175
0.32	0.169 0.143	0.156	0.091
0.16	0.111 0.113	0.112	0.047
0	0.062 0.068	0.065	--

Reactivity

Rat

Specificity

This kit recognizes Rat UCP-1 in samples. No significant cross-reactivity or interference between Rat UCP-1 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

0.156--10ng/mL

Linearity

Samples were spiked with high concentrations of Rat UCP-1 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 (%)	92-108	88-100	100-116
1:2	Average (%)	99	93	106
1:4	Range (%)	90-102	84-93	84-99
1:4	Average (%)	95	88	90
1:8	Range (%)	94-107	85-95	85-97
1:8	Average (%)	99	90	90
1:16	Range (%)	86-100	81-92	84-100
1:16	Average (%)	93	86	91

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat UCP-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat UCP-1 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)	0.50	1.44	3.83	0.54	1.53	4.11
Standard	deviation	0.03	0.07	0.13	0.04	0.06	0.21
C V (%)	6.00	4.86	3.39	7.41	3.92	5.11

Recovery

The recovery of Rat UCP-1 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)	91-107	99
EDTA plasma (n=5)	95-107	101
Cell culture media (n=5)	93-105	98

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

0.1 ng/mL

Uniprot	P04633

Manual

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SKU	Supplier No.	Size	Your price
BHE10307034	E-EL-R1146	96T	$609	Add to Cart
BHE10307034	E-EL-R1146	96T*5	$2740.50	Add to Cart
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BHE10307034	E-EL-R1146	48T	$487.20	Add to Cart

Rat UCP-1(Uncoupling Protein 1, Mitochondrial) ELISA Kit

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