EPO ELISA Kit (Mouse) (OKBB00140)

Catalog No: OKBB00140

Size:96 Tests

Mouse EPO ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Mouse
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's mouse EPO ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for EPO has been precoated onto 96-well plates. Standards(NS0, A27-R192)and test samples are added to the wells, a biotinylated detection monoclonal antibody from rat specific for EPO 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 EPO amount of sample captured in plate.
• ELISA Kit Range: 46.9 pg/ml - 3,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	225	457	1046	246	486	1118
  St. Dev.	10.13	17.4	55.4	23.12	36.45	43.6
  %CV	4.50	3.80	5.29	9.39	7.5	3.89

• ELISA Kit Component:
  • Lyophilized recombinant mouse EPO standard: 10 ng/tube x 2
  • Anti-mouse EPO Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse EPO antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 46.9pg/ml-3000pg/ml
  Principle: Aviva’s mouse EPO ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse EPO species-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 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 mouse EPO 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: Erythropoietin, EPO, also known as hematopoietin or hemopoietin, is a glycoprotein hormone that controls erythropoiesis, or red blood cell production. It is a cytokine for erythrocyte (red blood cell) precursors in the bone marrow. Its gene is mapped to 7q22. It is said that the EPO gene encodes a deduced 193-amino acid propolypeptide. This hormone can be found in kidney and liver. It is the hormone that regulates red blood cell production. And it plays an important role in the brain's response to neuronal injury. What’s more, EPO is also involved in the wound healing process.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum, plasma (heparin) and tissue homogenates
• Sensitivity: <15 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Siren AL et al. (2001). "Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress".
  Proc Natl Acad Sci USA 98 (7): 4044–4049. doi:10.1073/pnas.051606598. PMC 31176. PMID 11259643.
  2. Haroon ZA, Amin K, Jiang X, Arcasoy MO (September 2003). "A novel role for erythropoietin during fibrin-induced
  wound-healing response". Am. J. Pathol. 163 (3): 993–1000. PMC 1868246. PMID 12937140.
  3. Brines, M. L., Ghezzi, P., Keenan, S., Agnello, D., de Lanerolle, N. C., Cerami, C., Itri, L. M., Cerami, A.
  Erythropoietin crosses the blood-brain barrier to protect against experimental brain injury. Proc. Nat. Acad. Sci. 97:
  10526-10531, 2000. [PubMed: 10984541, related citations] [Full Text: HighWire Press, Pubget]
• Specificity: Natural and recombinant mouse EPO
• Immunogen: Expression system for standard: NS0; Immunogen sequence: A27-R192
• Description: For quantitative detection of mouse EPO in cell culture supernatants, serum and plasma(heparin).

Target Info

• Gene Symbol: EPO
• Gene Full Name: erythropoietin
• Alias Symbols: EP, EPO, EPO_HUMAN, Epoetin, Erythropoietin, MVCD2
• NCBI Gene Id: 13856
• Protein Name: Erythropoietin
• Description of Target: Erythropoietin, EPO, also known as hematopoietin or hemopoietin, is a glycoprotein hormone that controls erythropoiesis, or red blood cell production. It is a cytokine for erythrocyte (red blood cell) precursors in the bone marrow. Its gene is mapped to 7q22. It is said that the EPO gene encodes a deduced 193-amino acid propolypeptide. This hormone can be found in kidney and liver. It is the hormone that regulates red blood cell production. And it plays an important role in the brain's response to neuronal injury. What’s more, EPO is also involved in the wound healing process.
• Uniprot ID: P07321
• Protein Accession #: NP_031968.1
• Nucleotide Accession #: NM_007942.2