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Home » Products » Elisa Kits » Rat CASP8(Caspase 8) ELISA Kit

Rat CASP8(Caspase 8) ELISA Kit

Rat CASP8(Caspase 8) ELISA Kit
Rat CASP8(Caspase 8) ELISA Kit
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Product Code : E-EL-R0280
Brand Name : elabscience
Product Description

Rat CASP8(Caspase 8) ELISA Kit

Intended use

This ELISA kit applies to the in vitro quantitative determination of Rat CASP8 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 antibody specific to Rat CASP8. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat CASP8 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 CASP8, biotinylated detection antibody 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 CASP8. You can calculate the concentration of Rat CASP8 in the samples by comparing the OD of the samples to the standard curve.

Assay type

Sandwich

Format

96T

Assay time

4.5h

Reactivity

Rat

Detection Method

Colormetric

Detection Range

0.16 10 ng/mL

Sensitivity

0.10 ng/mL

Sample Volume Required Per Well

100 ¼L

Sample Type

Serum, plasma and other biological fluids

Specificity

This kit recognizes Rat CASP8 in samples. No significant cross-reactivity or interference between Rat CASP8 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

10

2.437
2.461

2.449

2.389

5

1.653
1.667

1.66

1.6

2.5

0.969
0.957

0.963

0.903

1.25

0.437
0.475

0.456

0.396

0.63

0.263
0.263

0.263

0.203

0.32

0.177
0.163

0.17

0.11

0.16

0.109
0.123

0.116

0.056

0

0.054
0.066

0.06

--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat CASP8 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat CASP8 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)

0.48

1.38

4.76

0.44

1.46

4.79

Standard deviation

0.03

0.08

0.18

0.02

0.07

0.18

C V (%)

6.25

5.80

3.78

4.55

4.79

3.76

Recovery


The recovery of Rat CASP8 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)

93-106

99

EDTA plasma (n=5)

92-103

97

Cell culture media (n=5)

87-98

92

Linearity

Samples were spiked with high concentrations of Rat CASP8 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 (%)

96-110

92-104

100-114

Average (%)

102

99

106

1:4

Range (%)

90-102

82-95

82-93

Average (%)

96

88

88

1:8

Range (%)

93-107

85-95

83-96

Average (%)

100

90

90

1:16

Range (%)

91-105

85-98

84-99

Average (%)

96

92

90


Kit components & Storage

An unopened kit can be stored at 4 ƒ 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„ƒ, 6 months

Reference Standard

2 vials

Concentrated Biotinylated Detection Ab (100 )

1 vial, 120 ¼L

Concentrated HRP Conjugate (100 )

1 vial, 120 ¼L

-20„ƒ(shading light), 6 months

Reference Standard & Sample Diluent

1 vial, 20 mL

4„ƒ, 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„ƒ(shading light)

Stop Solution

1 vial, 10 mL

4„ƒ

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




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