Nori Equine IL-29 ELISA Kit

$461.00$832.00

DataSheet   CoA   SDS

This ELISA kit is for quantification of IL-29 in horse. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for IL-29 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-29 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for IL-29 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of IL-29 bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for IL-29: IL29, interleukin 29, interferon lambda 1, IFNL1

This product is for Laboratory Research Use Only not for diagnostics or therapeutics or any other purposes.

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Description

Nori Equine IL-29 ELISA Kit Summary

Alternative names for IL-29: IL29, interleukin 29, interferon lambda 1, IFNL1

Alternative name for equine: Horse

Assay Type Solid Phase Sandwich ELISA
Format 96-well Microplate or 96-Well Strip Microplate
Method of Detection Colorimetric
Number of Targets Detected 1
Target Antigen Accession Number na
Assay Length 3 hours
Quantitative/Semiquantitative Quantitative
Sample Type Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL,
Recommended Sample Dilution (Plasma/Serum) No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ
Sensitivity 5 pg/mL
Detection Range 25-1600 pg/mL
Specificity Natural and recombinant equine IL-29
Cross-Reactivity < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested.
Interference No significant interference observed with available related molecules
Storage/Stability 4 ºC for up to 6 months
Usage For Laboratory Research Use Only. Not for diagnostic or therapeutic use.
Additional Notes The kit allows for use in multiple experiments.

 

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Interleukin-29 (IL-29) is a protein that is encoded by the IL29 gene.[1] IL-29 is a member of the helical cytokine family and is a type III interferon. It is also known as IFNλ1 and is highly similar in amino acid sequence to the IL-28, the other type III interferon. IL-29 plays an important role in host defenses against microbes and its gene is highly upregulated in cells infected with viruses. IL29 is not present in the mouse genome. IL28A, IL28B, and IL29, also named interferon λ2 (IFNλ2), IFNλ3, and IFNλ1, respectively, are class II cytokine receptor ligands that are distantly related to members of the IL10 family (11-13% aa sequence identity) and the type I IFN family (15-19% aa sequence identity). [2-4] The expression of IL-28A, IL-28B, and IL29 is induced by virus infection or double stranded RNA. All three cytokines exert bioactivities that overlap those of type I IFNs, including antiviral activity and upregulation of MHC class I antigen expression. The three proteins signal through the same heterodimeric receptor complex that is composed of the IL10 receptor β (IL10 Rβ) and a novel IL28 receptor α (IL28Rα, also known as IFNλ R1). Ligand binding to the receptor complex induces Jak kinase activation and STAT1 and STAT2 tyrosine phosphorylation. The phosphorylated STAT1 and STAT2 complex with IFN regulatory factor 9 (IRF9) to form the IFN stimulated regulatory factor 3 (ISGF3) transcription factor complex that is translocated to the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) present in the regulatory region of the target genes.

 

References

  1. Sheppard P, et al. (2003). Nature Immunology. 4(1): 63–8.
  2. Vilcek, J. (2003) Nature Immunol. 4:8.
  3. Sheppard, P. et al. (2003) Nature Immunol. 4:63.
  4. Kotenko, S.V. et al. (2003) Nature Immunol. 4:69.

 

 

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