Rat GFAP (Glial Fibrillary Acidic Protein) CLIA Kit-Laboratory Biological Research Reagents Manufacturer for Life Science - Elabscience

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Rat GFAP (Glial Fibrillary Acidic Protein) CLIA Kit

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Price:   $682
Cat Number:  E-CL-R0740
Lead Time:   Order now, ship in 3 days

Reactivity:  Rat

Detection Range:  62.50~4000 pg/mL

Sensitivity:  37.50 pg/mL

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Intended use

This CLIA kit applies to the in vitro quantitative determination of Rat GFAP concentrations in serum, plasma and other biological fluids.

Test principle

This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat GFAP. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat GFAP 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 GFAP, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat GFAP. You can calculate the concentration of Rat GFAP in the samples by comparing the RLU value of the samples to the standard curve.

Assay type Sandwich-CLIA
Format 96T
Assay time 3.5h
Reactivity Rat
Detection method Chemiluminescence
Detection range 62.50-4000 pg/mL
Sensitivity 37.50 pg/mL
Sample volume 100μL
Sample type serum, plasma and other biological fluids
Repeatability CV < 15%

Specificity

This kit recognizes Rat GFAP in samples. No significant cross-reactivity or interference between Rat GFAP and analogues was observed.

Typical data

As the RLU 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)
RLU Average Corrected
4000 57483
60679
59081 59046
2000 26499
29219
27859 27824
1000 13873
13095
13484 13449
500 6489
6721
6605 6570
250 3268
3218
3243 3208
125 1642
1520
1581 1546
62.50 724
786
755 720
0 35
35
35 --
CLIA-Elabscience

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat GFAP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat GFAP 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) 198.82 517.36 1456.27 214.62 541.21 1491.35
Standard deviation 16.22 43.46 156.26 26.27 59.70 141.68
C V (%) 8.16 8.40 10.73 12.24 11.03 9.50

Recovery

The recovery of Rat GFAP 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) 90-107 97
EDTA plasma (n=5) 84-100 91
Cell culture media (n=5) 101-117 108

Linearity

Samples were spiked with high concentrations of Rat GFAP 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 (%) 93-107 94-107 90-105
Average (%) 100 100 96
1:4 Range (%) 101-118 86-97 87-100
Average (%) 108 92 94
1:8 Range (%) 98-112 92-107 89-102
Average (%) 106 98 96
1:16 Range (%) 101-114 94-106 89-101
Average (%) 107 101 95

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 CLIA 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 A 1 vial, 5 mL 4℃ (shading light)
Substrate Reagent B 1 vial, 5 mL 4℃ (shading light)
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

  • Chemiluminescence immunoassay analyzer
  • 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 of standard or sample to each well. Incubate for 90 min at 37℃.

2.Remove the liquid.

3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times.

4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃. Aspirate and wash 5 times.

5.Add 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃.

6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results.

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