Rat CRP (C-Reactive Protein) CLIA Kit (RTES00018)

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SKU:
RTES00018
€649

Description

ELISA Kit Technical ManualMSDS

Rat CRP (C-Reactive Protein) 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 CRP . 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 CRP 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 CRP, 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 CRP. The concentration of Rat CRP 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 range0.16-10 ng/mL
Sensitivity0.09 ng/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat CRP in samples. No significant cross-reactivity or interference between Rat CRP 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
1031708
33234
3247132447
514599
16341
1547015446
2.57771
7235
75037479
1.253474
3832
36533629
0.631808
1714
17611737
0.31834
814
824800
0.16336
378
357333
024
24
24--

Precision

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

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

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.531.454.780.541.464.49
Standard deviation0.050.110.310.050.170.35
C V (%)9.437.596.499.2611.647.80

Recovery

The recovery of Rat CRP 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)99-115105
EDTA plasma (n=5)87-10193
Cell culture media (n=5)100-112107

Linearity

Samples were spiked with high concentrations of Rat CRP 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 (%)99-11496-11087-100
Average (%)10510294
1:4Range (%)103-11685-9692-105
Average (%)1099199
1:8Range (%)88-102102-11592-108
Average (%)95109100
1:16Range (%)94-11285-98100-113
Average (%)10290106

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 CRP (C-Reactive Protein) CLIA Kit (RTES00018) 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 CRP (C-Reactive Protein) CLIA Kit (RTES00018) Protein Information

UniProt Protein Function:Displays several functions associated with host defense: it promotes agglutination, bacterial capsular swelling, phagocytosis and complement fixation through its calcium-dependent binding to phosphorylcholine. Can interact with DNA and histones and may scavenge nuclear material released from damaged circulating cells.
NCBI Summary:glycoprotein of the acute phase response [RGD, Feb 2006]
UniProt Code:P48199
NCBI GenInfo Identifier:1345834
NCBI Gene ID:25419
NCBI Accession:P48199.1
UniProt Secondary Accession:P48199,Q5BK94,
UniProt Related Accession:P48199
Molecular Weight:25,468 Da
NCBI Full Name:C-reactive protein
NCBI Synonym Full Names:C-reactive protein, pentraxin-related
NCBI Official Symbol:Crp  
NCBI Official Synonym Symbols:Aa1249; Ac1262; Ab1-341; Ab2-196; Ac1-114; Ac2-069; Ba2-693  
NCBI Protein Information:C-reactive protein; C-reactive protein, petaxin related; C-reactive protein member of the pentraxin family; C-reactive protein, member of the pentraxin family
UniProt Protein Name:C-reactive protein
Protein Family:Cryparin
UniProt Gene Name:Crp  
UniProt Entry Name:CRP_RAT
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Additional Information

Reactivity:
Rat
ELISA Type:
CLIA
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