Rat PKIA(Protein Kinase Inhibitor Alpha) ELISA Kit

Mfr.No.

E-EL-R0818

Antigen

PKIA

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(pg/mL)	O.D	Average	Corrected
4000	2.427 2.461	2.444	2.373
2000	1.531 1.587	1.559	1.488
1000	0.889 0.875	0.882	0.8110000000000001
500	0.419 0.445	0.432	0.361
250	0.241 0.233	0.237	0.166
125	0.175 0.161	0.168	0.097
62.50	0.116 0.126	0.121	0.05
0	0.068 0.074	0.07099999999999999	--

Reactivity

Rat

Specificity

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

62.5--4000pg/mL

Linearity

Samples were spiked with high concentrations of Rat PKIA 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 (%)	95-112	89-101	93-110
1:2	Average (%)	102	94	100
1:4	Range (%)	90-101	83-97	86-97
1:4	Average (%)	96	89	91
1:8	Range (%)	92-105	85-99	85-96
1:8	Average (%)	98	92	90
1:16	Range (%)	87-101	85-99	87-99
1:16	Average (%)	94	90	93

Method Type

Sandwich

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat PKIA were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat PKIA 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)	213.38	415.61	1787.45	208.78	391.67	1898.58
Standard	deviation	11.44	22.19	71.50	13.49	21.35	93.79
C V (%)	5.36	5.34	4.00	6.46	5.45	4.94

Recovery

The recovery of Rat PKIA 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)	86-99	93
EDTA plasma (n=5)	89-102	96
Cell culture media (n=5)	92-103	97

Sample Type

Serum, Plasma, Cell supernatant

Sensitivity

37.5 pg/mL

Manual

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SKU	Supplier No.	Size	Your price
BHE10306790	E-EL-R0818	96T	$609	Add to Cart
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BHE10306790	E-EL-R0818	48T	$487.20	Add to Cart

Rat PKIA(Protein Kinase Inhibitor Alpha) ELISA Kit

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