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Rat HPF4-Ab(Anti-Heparin/Platelet Factor 4) ELISA Kit

Catalog number:E-EL-R2589

Synonyms:HPF4-Ab

Size:

96T
24T

Qty:

- +

Price: $580

Reactivity: Rat

Detection Range: 0.78~50 ng/mL

Sensitivity: 0.47 ng/mL

Lead Time: 7~10 daysWelcome to order from local distributors.

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Intended use

This ELISA kit applies to the in vitro quantitative determination of Rat HPF4-Ab concentrations in serum, plasma and other biological fluids.

Test principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antigen specific to Rat HPF4-Ab. Standards or samples are added to the micro ELISA plate wells and combined with the specific antigen. Then a biotinylated detection antigen specific for Rat HPF4-Ab and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat HPF4-Ab, biotinylated detection antigen and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat HPF4-Ab. You can calculate the concentration of Rat HPF4-Ab in the samples by comparing the OD of the samples to the standard curve.

Assay time	4.5h
Format	96T
Reactivity	Rat
Detection Method	Colormetric
Detection Range	0.78—50 ng/mL
Sensitivity	0.47 ng/mL
Sample Volume	100μL
Sample Type	Serum, plasma and other biological fluids

Specificity

This kit recognizes Rat HPF4-Ab in samples. No Significant cross-reactivity or interference between Rat HPF4-Ab and analogues was observed.

Typical data

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
50	2.492 2.516	2.504	2.424
25	1.641 1.645	1.643	1.563
12.5	0.954 0.92	0.937	0.857
6.25	0.487 0.509	0.498	0.418
3.13	0.269 0.267	0.268	0.188
1.57	0.178 0.178	0.178	0.098
0.78	0.121 0.139	0.13	0.05
0	0.075 0.085	0.08	--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat HPF4-Ab were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat HPF4-Ab 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)	2.57	4.39	19.76	2.72	4.18	19.67
Standard deviation	0.17	0.26	0.77	0.15	0.24	0.78
C V (%)	6.61	5.92	3.90	5.51	5.74	3.97

Recovery

The recovery of Rat HPF4-Ab spiked at two different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=5)	95-108	101
EDTA plasma (n=5)	87-98	93

Linearity

Samples were spiked with high concentrations of Rat HPF4-Ab 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)
1:2	Range (%)	89-100	89-101
1:2	Average (%)	95	95
1:4	Range (%)	84-97	86-99
1:4	Average (%)	90	92
1:8	Range (%)	85-101	83-95
1:8	Average (%)	92	88
1:16	Range (%)	83-95	82-92
1:16	Average (%)	89	87

Kit components & Storage

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 Ag (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 Ag 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

Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

Other supplies required

Microplate reader with 450 nm wavelength filter
High-precision transfer pipette, EP tubes and disposable pipette tips
Incubator capable of maintaining 37℃
Deionized or distilled water
Absorbent paper
Loading slot for Wash Buffer

Assay Procedure

	1.Add 100 μL standard or sample to each well. Incubate for 90 min at 37℃.
	2.Remove the liquid.
	3.Add 100 μL Biotinylated Detection Ag. Incubate for 1 hour at 37℃. Aspirate and wash 3 times.
	4.Add 100 μL HRP Conjugate. Incubate for 30 min at 37℃. Aspirate and wash 5 times.
	5.Add 90 μL of Substrate Reagent. Incubate for 15 min at 37℃.
	6.Add 50 μL Stop Solution.
	7.Read at 450 nm immediately. Calculation of results.