Mouse CXCL9/MIG(Monokine induced by interferon-gamma)CLIA Kit (MOES00018)

SKU:
MOES00018
€649

Description

ELISA Kit Technical ManualMSDS

Mouse CXCL9/MIG(Monokine induced by interferon-gamma)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 CXCL9/MIG . 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 CXCL9/MIG 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 CXCL9/MIG, 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 CXCL9/MIG. The concentration of Mouse CXCL9/MIG 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 range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Mouse CXCL9/MIG in samples. No significant cross-reactivity or interference between Mouse CXCL9/MIG 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
100051360
55990
5367553650
50021393
23585
2248922464
25010893
9529
1021110186
1254612
5190
49014876
62.52575
2331
24532428
31.251287
1275
12811256
15.63697
719
708683
025
25
25--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse CXCL9/MIG were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse CXCL9/MIG were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)48.54142.04469.7152.58129.98462.13
Standard deviation5.4910.1343.965.9614.1531.93
C V (%)11.317.139.3611.3410.896.91

Recovery

The recovery of Mouse CXCL9/MIG 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)83-9590
EDTA plasma (n=5)92-10798
Cell culture media (n=5)89-10194

Linearity

Samples were spiked with high concentrations of Mouse CXCL9/MIG 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 (%)99-11599-10985-98
Average (%)10710492
1:4Range (%)89-10193-108100-118
Average (%)94101108
1:8Range (%)97-112100-11387-102
Average (%)10410794
1:16Range (%)86-10095-11191-103
Average (%)9210297

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 CXCL9/MIG(Monokine induced by interferon-gamma)CLIA Kit (MOES00018) 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 CXCL9/MIG(Monokine induced by interferon-gamma)CLIA Kit (MOES00018) Protein Information

UniProt Protein Function:CXCL9: Cytokine that affects the growth, movement, or activation state of cells that participate in immune and inflammatory response. Chemotactic for activated T-cells. Binds to CXCR3. 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; external side of plasma membrane

Molecular Function:CXCR3 chemokine receptor binding; chemokine activity; cytokine activity

Biological Process: G-protein coupled receptor protein signaling pathway; positive regulation of leukocyte chemotaxis; immune response; response to lipopolysaccharide; chemotaxis; inflammatory response; defense response to virus; positive regulation of myoblast differentiation; regulation of cell proliferation

UniProt Code:P18340
NCBI GenInfo Identifier:408360042
NCBI Gene ID:17329
NCBI Accession:P18340.2
UniProt Secondary Accession:P18340,Q8C9J0, Q99J60,
UniProt Related Accession:P18340
Molecular Weight:14,444 Da
NCBI Full Name:C-X-C motif chemokine 9
NCBI Synonym Full Names:chemokine (C-X-C motif) ligand 9
NCBI Official Symbol:Cxcl9
NCBI Official Synonym Symbols:CMK; Mig; MuMIG; Scyb9; crg-10; BB139920
NCBI Protein Information:C-X-C motif chemokine 9; M119; protein m119; small-inducible cytokine B9; gamma interferon-induced monokine; gamma-interferon-induced monokine; monokine induced by gamma interferon; monokine induced by interferon-gamma; small inducible cytokine B subfamily (Cys-X-Cys), member 9
UniProt Protein Name:C-X-C motif chemokine 9
UniProt Synonym Protein Names:Gamma-interferon-induced monokine; Monokine induced by interferon-gamma; MIG; MuMIG; Protein m119; Small-inducible cytokine B9
Protein Family:Migration and invasion enhancer
UniProt Gene Name:Cxcl9
UniProt Entry Name:CXCL9_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
CLIA
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