Mouse GCP-2 (Granulocyte Chemotactic Protein 2) CLIA Kit (MOES00310)

SKU:
MOES00310
Product Type:
CLIA Kit
Size:
96 Assays
Sensitivity:
7.5pg/mL
Range:
12.5-800pg/mL
ELISA Type:
Sandwich
Synonyms:
CXCL6, GCP2, SCYB6, CKA-3
Reactivity:
Mouse
Tested Sample Types:
Serum, plasma and other biological fluids
€649

Description

ELISA Kit Technical ManualMSDS

Mouse GCP-2 (Granulocyte Chemotactic Protein 2) 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 Mouse GCP-2 . 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 Mouse GCP-2 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 Mouse GCP-2, 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 Mouse GCP-2. The concentration of Mouse GCP-2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection methodChemiluminescence
Detection range12.50-800 pg/mL
Sensitivity7.50 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Mouse GCP-2 in samples. No significant cross-reactivity or interference between Mouse GCP-2 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
80052571
52713
5264252616
40021670
21798
2173421708
20010063
9353
97089682
1004450
4654
45524526
502213
2163
21882162
251067
1053
10601034
12.50478
540
509483
025
27
26--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse GCP-2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse GCP-2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)39.1468.47348.7541.9362.40362.61
Standard deviation4.335.3538.474.347.1922.48
C V (%)11.067.8111.0310.3511.526.20

Recovery

The recovery of Mouse GCP-2 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)88-10193
EDTA plasma (n=5)91-10196
Cell culture media (n=5)86-9992

Linearity

Samples were spiked with high concentrations of Mouse GCP-2 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 (%)95-110101-117100-112
Average (%)100107107
1:4Range (%)97-11395-11294-108
Average (%)105102101
1:8Range (%)93-10984-9996-110
Average (%)10091102
1:16Range (%)85-9892-10591-106
Average (%)9210098

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

Mouse GCP-2 (Granulocyte Chemotactic Protein 2) CLIA Kit (MOES00310) 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 100 µL of standard solutions into the standard wells.
    3. Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 100 µL 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.

Mouse GCP-2 (Granulocyte Chemotactic Protein 2) CLIA Kit (MOES00310) Protein Information

UniProt Protein Function:CXCL6: Chemotactic for neutrophil granulocytes. Belongs to the intercrine alpha (chemokine CxC) family.
UniProt Protein Details:

Protein type:Secreted; Secreted, signal peptide; Motility/polarity/chemotaxis

Cellular Component: extracellular space; extracellular region

Molecular Function:chemokine activity; CXCR chemokine receptor binding; cytokine activity

Biological Process: G-protein coupled receptor protein signaling pathway; organ regeneration; cytokine production; positive regulation of JAK-STAT cascade; immune response; positive regulation of leukocyte chemotaxis; response to lipopolysaccharide; inflammatory response; chemotaxis; regulation of chemokine production; regulation of cell proliferation; leukocyte homeostasis

NCBI Summary:This gene encodes a protein that is a member of the CXC subfamily of chemokines. Chemokines, which recruit and activate leukocytes, are classified by function (inflammatory or homeostatic) or by structure. This protein is proposed to bind the G-protein coupled receptor chemokine (C-X-C motif) receptor 2 to recruit neutrophils and to have homeostatic and inflammatory functions. In mouse, deficiency of this gene is associated with increased lung inflammation that is neutrophil-dependent. [provided by RefSeq, May 2013]
UniProt Code:P50228
NCBI GenInfo Identifier:408360041
NCBI Gene ID:20311
NCBI Accession:P50228.2
UniProt Secondary Accession:P50228,Q8K3I3,
UniProt Related Accession:P50228
Molecular Weight:14,235 Da
NCBI Full Name:C-X-C motif chemokine 5
NCBI Synonym Full Names:chemokine (C-X-C motif) ligand 5
NCBI Official Symbol:Cxcl5
NCBI Official Synonym Symbols:LIX; GCP-2; Scyb5; Scyb6; ENA-78; AMCF-II
NCBI Protein Information:C-X-C motif chemokine 5; cytokine LIX; small-inducible cytokine B5; granulocyte chemotactic protein-2; small inducible cytokine B subfamily, member 5; small inducible cytokine subfamily B, member 15
UniProt Protein Name:C-X-C motif chemokine 5
UniProt Synonym Protein Names:Cytokine LIX; Small-inducible cytokine B5Cleaved into the following 2 chains:GCP-2(1-78); GCP-2(9-78)
UniProt Gene Name:Cxcl5
UniProt Entry Name:CXCL5_MOUSE