Rat GRO alpha/CXCL1 (Growth Regulated Oncogene Alpha) CLIA Kit (RTES00003)

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RTES00003
€649

Description

ELISA Kit Technical ManualMSDS

Rat GRO alpha/CXCL1 (Growth Regulated Oncogene 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 GRO alpha/CXCL1 . 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 GRO alpha/CXCL1 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 GRO alpha/CXCL1, 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 GRO alpha/CXCL1. The concentration of Rat GRO alpha/CXCL1 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 range7.81-500 pg/mL
Sensitivity4.69 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat GRO alpha/CXCL1 in samples. No significant cross-reactivity or interference between Rat GRO alpha/CXCL1 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
50046393
56681
5153751508
25020345
21651
2099820969
1259845
8947
93969367
62.54353
4671
45124483
31.252442
2156
22992270
15.631325
1175
12501221
7.81720
760
740711
028
30
29--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat GRO alpha/CXCL1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat GRO alpha/CXCL1 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)22.7965.44213.3021.1664.47209.54
Standard deviation1.986.4018.712.647.4923.76
C V (%)8.699.788.7712.4811.6211.34

Recovery

The recovery of Rat GRO alpha/CXCL1 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)84-9990
EDTA plasma (n=5)92-10699
Cell culture media (n=5)96-110103

Linearity

Samples were spiked with high concentrations of Rat GRO alpha/CXCL1 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 (%)99-11589-105101-114
Average (%)10796107
1:4Range (%)100-11290-10697-112
Average (%)10697104
1:8Range (%)91-10485-9698-115
Average (%)9990106
1:16Range (%)98-11485-9892-103
Average (%)1059198

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 GRO alpha/CXCL1 (Growth Regulated Oncogene Alpha) CLIA Kit (RTES00003) 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 GRO alpha/CXCL1 (Growth Regulated Oncogene Alpha) CLIA Kit (RTES00003) Protein Information

UniProt Protein Function:CXCL2: Produced by activated monocytes and neutrophils and expressed at sites of inflammation. Hematoregulatory chemokine, which, in vitro, suppresses hematopoietic progenitor cell proliferation. GRO-beta(5-73) shows a highly enhanced hematopoietic activity. Belongs to the intercrine alpha (chemokine CxC) family.
UniProt Protein Details:

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

Cellular Component: extracellular space

Molecular Function:growth factor activity; chemokine activity; CXCR chemokine receptor binding

Biological Process: neutrophil chemotaxis; response to glucocorticoid stimulus; response to amphetamine; positive regulation of potassium ion transport; response to lipopolysaccharide; positive regulation of leukocyte chemotaxis; response to estradiol stimulus; regulation of cell proliferation; G-protein coupled receptor protein signaling pathway; elevation of cytosolic calcium ion concentration; response to heat; response to molecule of bacterial origin; positive regulation of superoxide release; response to gamma radiation; immune response; acute inflammatory response

NCBI Summary:acts as a neutrophil chemoattractant; may play a role in acute phase inflammatory response [RGD, Feb 2006]
UniProt Code:P14095
NCBI GenInfo Identifier:1170021
NCBI Gene ID:81503
NCBI Accession:P14095.2
UniProt Related Accession:P14095
Molecular Weight:10,249 Da
NCBI Full Name:Growth-regulated alpha protein
NCBI Synonym Full Names:chemokine (C-X-C motif) ligand 1
NCBI Official Symbol:Cxcl1  
NCBI Official Synonym Symbols:Gro1; CINC-1  
NCBI Protein Information:growth-regulated alpha protein; C-X-C motif chemokine 1; cytokine-induced neutrophil chemoattractant 1; platelet-derived growth factor-inducible protein KC; chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha)
UniProt Protein Name:Growth-regulated alpha protein
UniProt Synonym Protein Names:C-X-C motif chemokine 1; Cytokine-induced neutrophil chemoattractant 1; CINC-1; Platelet-derived growth factor-inducible protein KC
Protein Family:Growth-regulated alpha protein
UniProt Gene Name:Cxcl1  
UniProt Entry Name:GROA_RAT
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Additional Information

Product type:
ELISA
Reactivity:
Rat
ELISA Type:
CLIA
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