Rat Fga (Fibrinogen Alpha) CLIA Kit (RTES00208)

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SKU:
RTES00208
€649

Description

ELISA Kit Technical ManualMSDS

Rat Fga (Fibrinogen Alpha) CLIA Kit

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

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range0.31-20 ng/mL
Sensitivity0.19 ng/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat Fga in samples. No significant cross-reactivity or interference between Rat Fga 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
(ng/mL)
RLU Average Corrected
2031800
36164
3398233950
1011646
13294
1247012438
55432
5068
52505218
2.52311
2737
25242492
1.251474
1292
13831351
0.63877
857
867835
0.31615
631
623591
031
33
32--

Precision

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

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat Fga were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.942.437.840.942.398.09
Standard deviation0.110.270.730.090.230.69
C V (%)11.7011.119.319.579.628.53

Recovery

The recovery of Rat Fga 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)87-10192
EDTA plasma (n=5)95-109102
Cell culture media (n=5)97-110102

Linearity

Samples were spiked with high concentrations of Rat Fga 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)
1:2Range (%)97-11283-97101-119
Average (%)10590109
1:4Range (%)99-11489-10489-103
Average (%)1049595
1:8Range (%)92-10697-10995-107
Average (%)97102101
1:16Range (%)103-11698-11496-114
Average (%)109105104

Kit Components & Storage

An unopened kit can be stored at 4'C 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 X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Rat Fga (Fibrinogen Alpha) CLIA Kit (RTES00208) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Rat Fga (Fibrinogen Alpha) CLIA Kit (RTES00208) Protein Information

UniProt Protein Function:FGA: Fibrinogen has a double Function: yielding monomers that polymerize into fibrin and acting as a cofactor in platelet aggregation
UniProt Protein Details:

Protein type:Secreted, signal peptide; Secreted

Cellular Component: cell cortex; cell surface; cytoplasm; external side of plasma membrane; extracellular space; fibrinogen complex; rough endoplasmic reticulum

Molecular Function:cell adhesion molecule binding; metal ion binding; protein binding, bridging; receptor binding; structural molecule activity

Biological Process: acute-phase response; adaptive immune response; blood coagulation; cell-matrix adhesion; cellular protein complex assembly; fibrinolysis; induction of bacterial agglutination; innate immune response; plasminogen activation; platelet activation; positive regulation of exocytosis; positive regulation of heterotypic cell-cell adhesion; positive regulation of protein secretion; positive regulation of vasoconstriction; protein complex assembly; protein polymerization; response to calcium ion; response to cycloheximide; response to estradiol stimulus; response to genistein; response to morphine; signal transduction

NCBI Summary:plays a role in blood coagulation; may be involved in liver regeneration [RGD, Feb 2006]
UniProt Code:P06399
NCBI GenInfo Identifier:1706800
NCBI Gene ID:361969
NCBI Accession:P06399.3
UniProt Related Accession:P06399
Molecular Weight:60,490 Da
NCBI Full Name:Fibrinogen alpha chain
NCBI Synonym Full Names:fibrinogen alpha chain
NCBI Official Symbol:Fga  
NCBI Official Synonym Symbols:Fba5e; Ac1873  
NCBI Protein Information:fibrinogen alpha chain
UniProt Protein Name:Fibrinogen alpha chain
Protein Family:Fibrinogen
UniProt Gene Name:Fga  
UniProt Entry Name:FIBA_RAT
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Additional Information

Reactivity:
Rat
ELISA Type:
CLIA
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