ErbB-2 ELISA Kit (Human) (OKBB00322)

Catalog No: OKBB00322

Size:96 Tests

Human ERBB2 ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human ErbB-2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A polyclonal antibody from goat specific for ErbB-2 has been precoated onto 96-well plates. Standards(NS0, T23-T652) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for ErbB-2 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 ErbB-2 amount of sample captured in plate.
• ELISA Kit Range: 62.5 pg/ml - 4,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	348	1057	2334	361	1049	2645
  St. Dev.	14.62	45.45	91.03	21.3	67.14	140.2
  %CV	4.20	4.29	3.90	5.90	6.40	5.30

• ELISA Kit Component:
  • Lyophilized recombinant mouse C-MET standard: 10 ng/tube x 2
  • Anti-mouse C-MET Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse C-MET 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: 62.5pg/ml-4000pg/ml
  Principle: Aviva’s Human ErbB-2 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human ErbB-2 specific-specific polyclonal antibodies were precoated onto 96-well plates. The human 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 Human ErbB-2 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: HER2/neu (also known as ErbB-2) stands for "Human Epidermal growth factor Receptor 2" and is a protein giving higher aggressiveness in breast cancers. It is a member of the ErbB protein family, more commonly known as the epidermal growth factor receptor family. HER2/neu has also been designated as CD340 (cluster of differentiation 340) and p185. It is encoded by the ERBB2 gene.HER2 is a cell membrane surface-bound receptor tyrosine kinase and is normally involved in the signal transduction pathways leading to cell growth and differentiation. It is encoded within the genome by HER2/neu, a known proto-oncogene. HER2 is thought to be an orphan receptor, with none of the EGF family of ligands able to activate it. However, ErbB receptors dimerise on ligand binding, and HER2 is the preferential dimerisation partner of other members of the ErbB family.1 The HER2 gene is a proto-oncogene located at the long arm of human chromosome 17(17q21-q22)2.
• 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) and tissue lysates
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1 . Olayioye MA (2001). "Update on HER-2 as a target for cancer therapy: intracellular signaling pathways of
  ErbB2/HER-2 and family members". Breast Cancer Res 3 (6): 385–389.
  2. Coussens L; Yang-Feng, TL; Liao, YC; Chen, E; Gray, A; McGrath, J; Seeburg, PH; Libermann, TA et al. (1985).
  "Tyrosine Kinase Receptor with Extensive Homology to EGF Receptor Shares Chromosomal Location with neu
  Oncogene". Science 230 (4730): 1132–1139.
• Specificity: Natural and recombinant human ErbB-2
• Immunogen: Expression system for standard: NS0; Immunogen sequence: T23-T652
• Description: For quantitative detection of human ErbB-2 in cell culture supernatants, cell lysates, serum and plasma(heparin, EDTA).

Target Info

• Gene Symbol: ERBB2
• Gene Full Name: v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2
• Alias Symbols: c erb B2, C erbB 2, CD340, CD340 antigen, Cerb B2/neu protein, CerbB2, Erb B2, erbb2, HER 2, HER 2/neu, HER2, Her2/neu, Herstatin, Metastatic lymph node gene 19 protein, MLN 19, MLN19, NEU, NEU Proto Oncogene, Neuro Glioblastoma Derived Oncogene Homolog, Neuroblastoma/glioblastoma derived oncogene homolog, NGL, p185 ErbB2, p185erbB2, Proto oncogene c ErbB 2, Proto oncogene Neu, Receptor protein tyrosine kinase ErbB2 precursor, Receptor tyrosine protein kinase erbB 2, TKR1, Tyrosine kinase type cell surface receptor HER2, v erb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog (avian), Verb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog
• NCBI Gene Id: 2064
• Protein Name: Receptor tyrosine-protein kinase erbB-2
• Description of Target: HER2/neu (also known as ErbB-2) stands for "Human Epidermal growth factor Receptor 2" and is a protein giving higher aggressiveness in breast cancers. It is a member of the ErbB protein family, more commonly known as the epidermal growth factor receptor family. HER2/neu has also been designated as CD340 (cluster of differentiation 340) and p185. It is encoded by the ERBB2 gene.HER2 is a cell membrane surface-bound receptor tyrosine kinase and is normally involved in the signal transduction pathways leading to cell growth and differentiation. It is encoded within the genome by HER2/neu, a known proto-oncogene. HER2 is thought to be an orphan receptor, with none of the EGF family of ligands able to activate it. However, ErbB receptors dimerise on ligand binding, and HER2 is the preferential dimerisation partner of other members of the ErbB family.1 The HER2 gene is a proto-oncogene located at the long arm of human chromosome 17(17q21-q22)2.
• Uniprot ID: P04626
• Protein Accession #: NP_001005862.1
• Nucleotide Accession #: NM_001005862.2