VCAM-1 ELISA Kit (Mouse) (OKBB00270)

Catalog No: OKBB00270

Size:96 Tests

Mouse VCAM1 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 VCAM-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for VCAM-1 has been precoated onto 96-well plates. Standards (NS0,F25 - E698) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for VCAM-1 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 VCAM-1 amount of sample captured in plate.
• ELISA Kit Range: 156 pg/ml - 10,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	230	2346	5393	384	3017	6245
  St. Dev.	8.74	105.6	280.4	17.28	162.9	374.7
  %CV	3.8	4.50	5.19	4.5	5.39	6

• ELISA Kit Component:
  • Lyophilized recombinant mouse VCAM1 standard: 10 ng/tube x 2
  • Anti-mouse VCAM1 Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse VCAM1 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: 156pg/ml-10,000pg/ml
  Principle: Aviva’s mouse VCAM-1 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse VCAM-1 specific-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 VCAM-1 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: Vascular cell adhesion molecule 1 (VCAM-1) is a cell surface glycoprotein adhesive for certain blood leukocytes and tumor cells, which is expressed by activated endothelium in a variety of pathologic conditions including atherosclerosis.Increased expression of VCAM1 is associated with a variety of chronic inflammatory conditions, making its expression and function a target for therapeutic intervention.Integrin alpha4beta1 (VLA-4) and VCAM-1 facilitate a critical cell-cell adhesion event required for survival of endothelial and mural cells during vascularization.The VCAM1 gene is assigned to the 1p31-32 region of chromosome 1 based on the analysis of human-mouse hybrid cell lines and in situ hybridization.The standard product used in this kit is recombinant mouse VCAM-1, a dimeric protein linking with a disulfide bond. It consists of Phe25-Glu698 amino acid sequence of a single chain with the molecular mass of 101.9KDa. As a result of glycosylation, the molecular mass of 120KDa is revealed by SDS-PAGE.
• 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 and plasma (heparin, EDTA)
• Sensitivity: <5 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Reference: 1. Cybulsky, M. I.; Fries, J. W. U.; Williams, A. J.; Sultan, P.; Eddy, R.; Byers, M.; Shows, T.; Gimbrone, M. A., Jr.;
  Collins, T. Gene structure, chromosomal location, and basis for alternative mRNA splicing of the human VCAM1
  gene. Proc. Nat. Acad. Sci. 88: 7859-7863, 1991.
  2. Besemer, J.; Harant, H.; Wang, S.; Oberhauser, B.; Marquardt, K.; Foster, C. A.; Schreiner, E. P.; de Vries, J. E.;
  Dascher-Nadel, C.; Lindley, I. J. D. Selective inhibition of cotranslational translocation of vascular cell adhesion
  molecule 1. (Letter) Nature 436: 290-293, 2005.
  3. Garmy-Susini, B.; Jin, H.; Zhu, Y.; Sung, R.-J.; Hwang, R.; Varner, J. Integrin alpha-4-beta-1--VCAM-1--mediated
  adhesion between endothelial and mural cells is required for blood vessel maturation. J. Clin. Invest. 115:
  1542-1551, 2005.
• Specificity: Natural and recombinant mouse VCAM-1
• Immunogen: Expression system for standard: NS0; Immunogen sequence: F25 - E698
• Description: For quantitative detection of mouse VCAM-1 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Target Info

• Gene Symbol: VCAM1
• Gene Full Name: vascular cell adhesion molecule 1
• Alias Symbols: CD106, CD106 antigen, DKFZp779G2333, INCAM 100, INCAM-100, INCAM100, L1CAM, MGC99561, V CAM 1, V-CAM 1, Vascular cell adhesion molecule 1, Vascular cell adhesion protein 1, VCAM 1, VCAM-1, VCAM1, VCAM1_HUMAN
• NCBI Gene Id: 22329
• Protein Name: Vascular cell adhesion protein 1
• Description of Target: Vascular cell adhesion molecule 1 (VCAM-1) is a cell surface glycoprotein adhesive for certain blood leukocytes and tumor cells, which is expressed by activated endothelium in a variety of pathologic conditions including atherosclerosis.Increased expression of VCAM1 is associated with a variety of chronic inflammatory conditions, making its expression and function a target for therapeutic intervention.Integrin alpha4beta1 (VLA-4) and VCAM-1 facilitate a critical cell-cell adhesion event required for survival of endothelial and mural cells during vascularization.The VCAM1 gene is assigned to the 1p31-32 region of chromosome 1 based on the analysis of human-mouse hybrid cell lines and in situ hybridization.The standard product used in this kit is recombinant mouse VCAM-1, a dimeric protein linking with a disulfide bond. It consists of Phe25-Glu698 amino acid sequence of a single chain with the molecular mass of 101.9KDa. As a result of glycosylation, the molecular mass of 120KDa is revealed by SDS-PAGE.
• Uniprot ID: P29533
• Protein Accession #: NP_035823.3
• Nucleotide Accession #: NM_011693.3