Fractalkine/CX3CL1 ELISA Kit (Mouse) (OKBB00149)

Catalog No: OKBB00149

Size:96 Tests

Mouse CX3CL1 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 Fractalkine ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for Fractalkine has been precoated onto 96-well plates. Standards(NS0, Q25-R337) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Fractalkine 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 Fractalkine amount of sample captured in plate.
• ELISA Kit Range: 312 pg/ml - 20,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (ng/mL)	1.57	3.5	6.3	1.32	3.7	6.4
  St. Dev.	0.113	0.224	0.473	0.11	0.285	0.544
  %CV	7.19	6.4	7.50	8.33	7.70	8.5

• ELISA Kit Component:
  • Lyophilized recombinant mouse Fractalkine standard: 10 ng/tube x 2
  • Anti-mouse Fractalkine Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-mouse Fractalkine 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 Fractalkine ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Mouse Fractalkine 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 Fractalkine 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: Chemokine (C-X3-C motif) ligand 1 (CX3CL1) is a large cytokine protein of 373 amino acids, it contains multiple domains and is the only known member of the CX3C chemokine family. It is also commonly known under the names fractalkine (in humans) and neurotactin (in mice).1, 2 CX3CL1 is produced as a long protein (with 373-amino acid in humans) with an extended mucin-like stalk and a chemokine domain on top. The mucin-like stalk permits it to bind to the surface of certain cells. However a soluble (90 kD) version of this chemokine has also been observed. Soluble CX3CL1 potently chemoattracts T cells and monocytes, while the cell-bound chemokine promotes strong adhesion of leukocytes to activated endothelial cells, where it is primarily expressed.2 CX3CL1 elicits its adhesive and migratory functions by interacting with the chemokine receptor CX3CR1.3 Its gene is located on human chromosome 16 along with some CC chemokines known as CCL17 and CCL22.2, 4 It can act as a mediator of smooth muscle cell migration in human atherosclerosis, rather than mediate inflammatory cell recruitment.5 The standard product used in this kit is recombinant mouse CX3CL1, consisting of 84 amino acids with the molecular mass of 9.5kDa.
• 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 homogenates
• Sensitivity: <10 pg/ml
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with Eotaxin, MCP-1, MCP-2, 6Ckine, MARC.
• Reference: 1. Pan et al. Neurotactin, a membrane-anchored chemokine upregulated in brain inflammation. Nature 387: 611-617,
  1997.
  2. Bazan et al. A new class of membrane-bound chemokine with a CX3C motif. Nature 385: 640-644, 1997.
  3. Imai et al. Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both
  leukocyte migration and adhesion. Cell 91: 521-530, 1997.
  4. Nomiyama et al. Human chemokines fractalkine (SCYD1), MDC (SCYA22) and TARC (SCYA17) are clustered on
  chromosome 16q13. Cytogenet. Cell Genet. 81: 10-11, 1998.
  5. Lucas, A. D.; Bursill, C.; Guzik, T. J.; Sadowski, J.; Channon, K. M.; Greaves, D. R. : Smooth muscle cells in human
  atherosclerotic plaques express the fractalkine receptor CX3CR1 and undergo chemotaxis to the CX3C chemokine
• Specificity: Natural and recombinant mouse Fractalkine
• Immunogen: Expression system for standard: NS0; Immunogen sequence: Q25-R337
• Description: For quantitative detection of mouse Fractalkine in cell culture supernatants, serum and plasma(heparin, EDTA).

Target Info

• Gene Symbol: CX3CL1
• Gene Full Name: chemokine (C-X3-C motif) ligand 1
• Alias Symbols: A 152E5.2, AB030188, ABCD 3, ABCD3, C X3 C motif chemokine 1, C3Xkine, Chemokine (C X3 C motif) ligand 1, Chemokine C X3 C motif ligand 1, Chemokine CX3C Motif Ligand 1, CX3C membrane anchored chemokine, CXC3, CXC3C, FKN, Fractalkine, Neurotactin, NTN, NTT, SCYD 1, SCYD1, Small inducible cytokine D1, Small inducible cytokine subfamily D (Cys X3 Cys), member 1, Small inducible cytokine subfamily D (Cys-X3-Cys), member 1 (fractalkine, neurotactin), Small inducible cytokine subfamily D member 1
• NCBI Gene Id: 6376
• Protein Name: Fractalkine
• Description of Target: Chemokine(C-X3-C motif) ligand 1(CX3CL1) is a large cytokine protein of 373 amino acids, it contains multiple domains and is the only known member of the CX3C chemokine family. It is also commonly known under the names fractalkine(in humans) and neurotactin(in mice). CX3CL1 is produced as a long protein(with 373-amino acid in humans) with an extended mucin-like stalk and a chemokine domain on top. The mucin-like stalk permits it to bind to the surface of certain cells. However a soluble(90 kD) version of this chemokine has also been observed. Soluble CX3CL1 potently chemoattracts T cells and monocytes, while the cell-bound chemokine promotes strong adhesion of leukocytes to activated endothelial cells, where it is primarily expressed. CX3CL1 elicits its adhesive and migratory functions by interacting with the chemokine receptor CX3CR1. Its gene is located on human chromosome 16 along with some CC chemokines known as CCL17 and CCL22. It can act as a mediator of smooth muscle cell migration in human atherosclerosis, rather than mediate inflammatory cell recruitment.
• Uniprot ID: O35188
• Protein Accession #: NP_033168.2
• Nucleotide Accession #: NM_009142.3