Human Endostatin ELISA

RayBio® Human Endostatin ELISA Kit for cell culture supernatants, plasma, and serum samples.

ELH-Endostatin-1
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Antigen Information

Accession Number:
  • P39060
Gene ID:
  • 80781
Gene Symbols:
  • COL18A1
Protein Name & Synonyms:
Collagen alpha-1(XVIII) chain [Cleaved into: Endostatin]
Target Species:
  • Human

Assay Format

Detection Range:
10 pg/ml - 10000 pg/ml
Recommended Dilution (serum & plasma):
100 fold
Sensitivity:
10 pg/ml

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Specificity:
This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
  1. Etulain J., Mena H., Negrotto S., Schattner M. Stimulation of PAR-1 or PAR-4 promotes similar pattern of VEGF and endostatin release and pro-angiogenic responses mediated by human platelets. Platelets. 2015;26(8):799-804. doi: 10.3109/09537104.2015.1051953.
    Species: Human
    Sample type: Platelet Releasate
  2. Biswal MR., Prentice HM., Dorey CK., Blanks JC. A hypoxia-responsive glial cell-specific gene therapy vector for targeting retinal neovascularization. Invest Ophthalmol Vis Sci. 2014 Nov 6;55(12):8044-53. doi: 10.1167/iovs.14-13932
    Species: Human
    Sample type: Tissue Lysate
  3. Etulain J., Negrotto S., Tribulatti M., Croci D., Carabelli J., et al. Control of Angiogenesis by Galectins Involves the Release of Platelet-Derived Proangiogenic Factors. PLOS One, Published: April 30, 2014 DOI: 10.1371/journal.pone.0096402 [Epub Ahead of Print]
    Species: Human
    Sample type: Conditioned Media
  4. Mohsen M., et al. Angiogenic output in viral hepatitis, C and B, and HCV-associated hepatocellular carcinoma . Alexandria Journal of Medicine Available online 4 July 2014 In Press, Corrected Proof. DOI: 10.1016/j.ajme.2014.06.003
    Species: Human
    Sample type: Serum
  5. Zhang Y., et al. Prognostic Values of VEGF and Endostatin with Malignant Pleural Effusions in Patients with Lung Cancer. Asian Pac J Cancer Prev. 2014;15(19):8435-40
    Species: Human
    Sample type: Serum
  6. Etulain J, Fondevila C, Negrotto S, Schattner M. Platelet-mediated angiogenesis is independent of VEGF and fully inhibited by aspirin. Br. J. Pharmacol. 2013 May 28. doi: 10.1111/bph.12250. [Epub ahead of print]
    Species: Human
    Sample type: Platelet Releasate
  7. Fujii, Masataka, et al. "Chondromodulin?I derived from the inner meniscus prevents endothelial cell proliferation." Journal of Orthopaedic Research 31.4 (2013): 538-543.
    Species: Human
    Sample type: Conditioned Media
  8. Jing, Yawu, et al. "Inhibition of ovarian cancer by RGD?P125A?endostatin?Fc fusion proteins." International journal of cancer 129.3 (2011): 751-761.
    Species: Human
    Sample type: Serum
  9. Etulain, J., et al. "Hyperthermia inhibits platelet hemostatic functions and selectively regulates the release of alpha?granule proteins." Journal of Thrombosis and Haemostasis 9.8 (2011): 1562-1571.
    Species: Human
    Sample type: Conditioned Media

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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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