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Catalog No: OKBB00256
Size:96 Tests
Price: $583.00
SKU
OKBB00256
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 ReactivityMouse
ApplicationELISA
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3 Hours
ELISA Kit PrincipleAviva's mouse TPO ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for TPO has been precoated onto 96-well plates. Standards(NS0, S22-T356) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TPO 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 mouse TPO amount of sample captured in plate.
ELISA Kit Range31.2 pg/ml - 2,000 pg/ml
ELISA Kit Reproducibility
Intra-Assay PrecisionInter-Assay Precision
Sample123123
Mean (pg/mL)29661112653217461464
St. Dev.11.843382.2316.747105.4
%CV45.406.505.206.307.19
ELISA Kit Component
ComponentAmount
Lyophilized recombinant mouse TPO standard10 ng/tube x 2
Anti-mouse TPO Antibody Well Plate96 Wells
Sample diluent buffer30 mL
Biotinylated anti-mouse TPO 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: 31.2pg/ml-2000pg/ml
::Principle: Aviva’s mouse TPO ELISA Kit was based on standard sandwich
enzyme-linked immune-sorbent assay technology. Mouse TPO specific
polyclonal antibodies were precoated onto 96-well plates. The mouse
specific detection polyclonal antibodies were biotinylated. The test samples
and biotinylated detection antibodies were added to the wells subsequently
and then followed by washing with TBS buffer. Avidin-Biotin-Peroxidase
Complex was added and unbound conjugates were washed away with 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 mouse TPO 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: Thrombopoietin (TPO) is also called megakaryocyte growth and development factor (MGDF). TPO, a lineage-specific cytokine affecting the proliferation and maturation of megakaryocytes from committed progenitor cells, is believed to be the major physiological regulator of circulating platelet levels.1 Human, dog, and mouse cDNAs for MGDF are highly conserved and encode open reading frames for proteins of 353, 352, and 356 amino acids, respectively, including predicted signal peptides. MGDF is a novel cytokine that regulates megakaryocyte development and is a ligand for the MPL receptor. The human MGDF gene has been mapped to chromosome 3q26.3. 2,3 The standard product used in this kit is gene recombinant mouse TPO, consisting of 335 amino acids with the molecular mass of 35KDa. As a result of glycosylation, the molecular mass is 75KDa.
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)
Sensitivity<5 pg/ml
Predicted Homology Based on Immunogen SequenceNo detectable cross-reactivity with any other cytokine.
Reference1. Foster, D. C.; Sprecher, C. A.; Grant, F. J.; Kramer, J. M.; Kuijper, J. L.; Holly, R. D.; Whitmore, T. E.; Heipel, M. D.;
Bell, L. A.; Ching, A. F. T.; McGrane, V.; Hart, C.; O'Hara, P. J.; Lok, S. Human thrombopoietin: gene structure,
cDNA sequence, expression, and chromosomal localization. Proc. Nat. Acad. Sci. 91: 13023-13027, 1994.
2. Bartley, T. D.; Bogenberger, J.; Hunt, P.; Li, Y.-S.; Lu, H. S.; Martin, F.; Chang, M.-S.; Samal, B.; Nichol, J. L.; Swift,
S.; Johnson, M. J.; Hsu, R.-Y.; and 41 others. Identification and cloning of a megakaryocyte growth and
development factor that is a ligand for the cytokine receptor Mpl. Cell 77: 1117-1124, 1994.
3. Chang, M.; McNinch, J.; Basu, R.; Shutter, J.; Hsu, R.; Perkins, C.; Mar, V.; Suggs, S.; Welcher, A.; Li, L.; Lu, H.;
Bartley, T.; Hunt, P.; Martin, F.; Samal, B.; Bogenberger, J. Cloning and characterization of the human
megakaryocyte growth and development factor (MGDF) gene. J. Biol. Chem. 270: 511-514, 1995.
SpecificityNatural and recombinant mouse TPO
ImmunogenExpression system for standard: NS0; Immunogen sequence: S22-T356
DescriptionFor quantitative detection of mouse TPO in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Gene SymbolTHPO
Gene Full Namethrombopoietin
Alias SymbolsC mpl ligand, Megakaryocyte colony stimulating factor, Megakaryocyte growth and development factor, Megakaryocyte stimulating factor, MGC163194, MGDF, MKCSF, ML, MPL ligand, MPLLG, Myeloproliferative leukemia virus oncogene ligand, OTTHUMP00000210504, THPO, TPO
NCBI Gene Id21832
Protein NameThrombopoietin
Description of TargetThrombopoietin (TPO) is also called megakaryocyte growth and development factor (MGDF). TPO, a lineage-specific cytokine affecting the proliferation and maturation of megakaryocytes from committed progenitor cells, is believed to be the major physiological regulator of circulating platelet levels.1 Human, dog, and mouse cDNAs for MGDF are highly conserved and encode open reading frames for proteins of 353, 352, and 356 amino acids, respectively, including predicted signal peptides. MGDF is a novel cytokine that regulates megakaryocyte development and is a ligand for the MPL receptor. The human MGDF gene has been mapped to chromosome 3q26.3. 2,3 The standard product used in this kit is gene recombinant mouse TPO, consisting of 335 amino acids with the molecular mass of 35KDa. As a result of glycosylation, the molecular mass is 75KDa.
Uniprot IDP40226
Protein Accession #NP_033443.1
Nucleotide Accession #NM_009417.3
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