EGF ELISA Kit (Mouse) (OKBB00136)

Catalog No: OKBB00136

Size:96 Tests

Mouse EGF 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 EGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for EGF has been precoated onto 96-well plates. Standards(E.coli, N977-R1029) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for EGF 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 EGF amount of sample captured in plate.
• ELISA Kit Range: 7.8 pg/ml - 500 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	11	42.5	186	9.8	47	192
  St. Dev.	0.539	2.252	9.858	0.049	2.867	11.136
  %CV	4.9	5.29	5.3	0.5	6.1	5.8

• ELISA Kit Component:
  • Lyophilized recombinant mouse EGF standard: 10 ng/tube x 2
  • Anti-mouse EGF Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse EGF 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: 15.6pg/ml-1000pg/ml
  Principle: Aviva’s mouse EGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse EGF specific-specific monoclonal 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 EGF 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: Epidermal growth factor or EGF is a growth factor that plays an important role in the regulation of cell growth, proliferation, and differentiation by binding to its receptor EGFR. EGF acts by binding with high affinity to epidermal growth factor receptor (EGFR) on the cell surface and stimulating the intrinsic protein-tyrosine kinase activity of the receptor. EGF results in cellular proliferation, differentiation, and survival.1 It also has a profound effect on the differentiation of specific cells in vivo and is a potent mitogenic factor for a variety of cultured cells of both ectodermal and mesodermal origin.2 EGF has strong expression in kidney, salivary gland, cerebrum, and prostate, moderate expression in trachea and thyroid, and low expression in bone marrow, heart, spleen, thymus, uterus, and colon. No expression was detected in adrenal gland, liver, lung, cerebellum, placenta, and small intestine.3
• 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, EDTA), tissue homogenates and urine
• Sensitivity: <1 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with other cytokine.
• Reference: 1. Herbst RS (2004). "Review of epidermal growth factor receptor biology". International Journal of Radiation Oncology,
  Biology, Physics 59 (2 Suppl): 21–6.
  2. Carpenter, G.; Cohen, S.: Epidermal growth factor. Ann. Rev. Biochem. 48: 193-216, 1979.
  3. Groenestege, W. M. T.; Thebault, S.; van der Wijst, J.; van den Berg, D.; Janssen, R.; Tejpar, S.; van den Heuvel, L.
  P.; van Cutsem, E.; Hoenderop, J. G.; Knoers, N. V.; Bindels, R. J. : Impaired basolateral sorting of pro-EGF causes
  isolated recessive renal hypomagnesemia. J. Clin. Invest. 117: 2260-2267, 2007.
• Specificity: Natural and recombinant mouse EGF
• Immunogen: Expression system for standard: E.coli; Immunogen sequence: N977-R1029
• Description: For quantitative detection of mouse EGF in cell culture supernatants, serum, plasma(heparin, EDTA)and urine.

Target Info

• Gene Symbol: EGF
• Gene Full Name: epidermal growth factor
• Alias Symbols: AI790464, Beta urogastrone, Epidermal growth factor, HOMG4, OTTHUMP00000219721, OTTHUMP00000219722, Pro epidermal growth factor, Pro epidermal growth factor precursor (EGF), URG, Urogastrone
• NCBI Gene Id: 13645
• Protein Name: Pro-epidermal growth factor
• Description of Target: Epidermal growth factor or EGF is a growth factor that plays an important role in the regulation of cell growth, proliferation, and differentiation by binding to its receptor EGFR. EGF acts by binding with high affinity to epidermal growth factor receptor (EGFR) on the cell surface and stimulating the intrinsic protein-tyrosine kinase activity of the receptor. EGF results in cellular proliferation, differentiation, and survival.1 It also has a profound effect on the differentiation of specific cells in vivo and is a potent mitogenic factor for a variety of cultured cells of both ectodermal and mesodermal origin.2 EGF has strong expression in kidney, salivary gland, cerebrum, and prostate, moderate expression in trachea and thyroid, and low expression in bone marrow, heart, spleen, thymus, uterus, and colon. No expression was detected in adrenal gland, liver, lung, cerebellum, placenta, and small intestine.3
• Uniprot ID: P01132
• Protein Accession #: NP_034243.2
• Nucleotide Accession #: NM_010113.4