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Catalog No: OKBB00187
Size:96 Tests
Price: $583.00
SKU
OKBB00187
Availability: Domestic: within 1-2 week delivery | International: within 1-2 week delivery
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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivityHuman
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva's human IL-5 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for IL-5 has been precoated onto 96-well plates. Standards(NS0, I20-S134) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for IL-5 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human IL-5 amount of sample captured in plate.
ELISA Kit Range7.8 pg/ml - 500 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)44.616833843.8188329
St. Dev.2.326.8914.872.6310.3421.06
%CV5.204.104.396.005.56.40
ELISA Kit Component
ComponentAmount
Lyophilized recombinant human IL-5 standard10 ng/tube x 2
Anti-human IL-5 Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-human IL-5 antibody130 uL, dilution 1:100
Antibody diluent buffer12 mL
Avidin-Biotin-Peroxidase Complex (ABC)130 uL, dilution 1:100
ABC diluent buffer12 mL
TMB color developing agent10 mL
TMB stop solution10 mL
Additional InformationRange: 15.6pg/ml-1000pg/ml
::Principle: Aviva’s human IL-5 ELISA Kit was based on standard sandwich
enzyme-linked immune-sorbent assay technology. Human IL-5 specific-specific
monoclonal antibodies were precoated onto 96-well plates. The human specific
detection monoclonal antibodies were biotinylated. The test samples and
biotinylated detection antibodies were added to the wells subsequently and
then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase
Complex was added and unbound conjugates were washed away with PBS or
TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction.
TMB was catalyzed by HRP to produce a blue color product that changed into
yellow after adding acidic stop solution. The density of yellow is proportional to
the human IL-5 amount of sample captured in plate.
::Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
experiment using standards and a small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
using.
Background: Interleukin-5 (IL-5) is also known as eosinophil differentiation factor (EDF). IL-5 is a potential candidate gene in the pathogenesis of asthma, as it is the main cytokine controlling eosinophil activity and eosinophils are pivotal in the development of airway inflammation.1 The predicted amino acid sequence of 134 amino acids is identical with that recently reported for human interleukin-5 but shows no significant homology with other known hemopoietic growth regulators.2 IL-5 is a lineage-specific hematopoietic growth factor that stimulates the production of eosinophils and eosinophil colonies from normal human bone marrow cells. IL-5 gene to human chromosome 5 at bands q23-31.3 Human and mouse of IL-5 share 70% amino acid sequence homology.
::1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot experiment using standards and a
small number of samples is recommended.
2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
4. Duplicate well assay is recommended for both standard and sample testing.
5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
6. Don’t reuse tips and tubes to avoid cross contamination.
7. To avoid to use the reagents from different batches together.
8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the marginal wells), it is
suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before using.
Reconstitution and StorageStore at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
Sample Typecell culture supernates, serum and plasma (heparin, EDTA) and urine
Sensitivity<2 pg/ml
Predicted Homology Based on Immunogen SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. Pereira, E.; Goldblatt, J.; Rye, P.; Sanderson, C.; Le Souef, P. Mutation analysis of interleukin-5 in an asthmatic
cohort. Hum. Mutat. 11: 51-54, 1998.
2. Campbell, H. D.; Tucker, W. Q. J.; Hort, Y.; Martinson, M. E.; Mayo, G.; Clutterbuck, E. J.; Sanderson, C. J.; Young, I.
G. Molecular cloning, nucleotide sequence, and expression of the gene encoding human eosinophil differentiation
factor (interleukin-5). Proc. Nat. Acad. Sci. 84: 6629-6633, 1987.
3. Le Beau, M. M.; Lemons, R. S.; Espinosa, R., III; Larson, R. A.; Arai, N.; Rowley, J. D. Interleukin-4 and interleukin-5
map to human chromosome 5 in a region encoding growth factors and receptors and are deleted in myeloid
leukemias with a del(5q). Blood 73: 647-650, 1989.
SpecificityNatural and recombinant human IL-5
ImmunogenExpression system for standard: NS0; Immunogen sequence: I20-S134
DescriptionFor quantitative detection of human IL-5 in cell culture supernatants, serum, plasma (heparin, EDTA) and urine.
Gene SymbolIL5
Gene Full Nameinterleukin 5
Alias SymbolsB cell differentiation factor I, B-cell differentiation factor I, Colony stimulating factor eosinophil, EDF, Eosinophil differentiation factor, IL 5, IL-5, IL5, IL5_HUMAN, Interleukin 5, Interleukin 5 (colony stimulating factor eosinophil), Interleukin-5, Interleukin5, T cell replacing factor, T-cell replacing factor, TRF
NCBI Gene Id3567
Protein NameInterleukin-5
Description of TargetInterleukin-5 (IL-5) is also known as eosinophil differentiation factor (EDF). IL-5 is a potential candidate gene in the pathogenesis of asthma, as it is the main cytokine controlling eosinophil activity and eosinophils are pivotal in the development of airway inflammation.1 The predicted amino acid sequence of 134 amino acids is identical with that recently reported for human interleukin-5 but shows no significant homology with other known hemopoietic growth regulators.2 IL-5 is a lineage-specific hematopoietic growth factor that stimulates the production of eosinophils and eosinophil colonies from normal human bone marrow cells. IL-5 gene to human chromosome 5 at bands q23-31.3 Human and mouse of IL-5 share 70% amino acid sequence homology.
Uniprot IDP05113
Protein Accession #NP_000870.1
Nucleotide Accession #NM_000879.2
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