Endostatin ELISA Assay Kit


The Endostatin ELISA Assay KIT(enzyme-linked immunoassay kit) is intended for the quantitative determination of human Endostatin in plasma and serum. The Eagle Biosciences Endostatin ELISA Assay KIT is for research use only and not to be used in diagnostic procedures.

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Endostatin ELISA Assay Kit

Endostatin ELISA Assay Kit Developed and Manufactured in Austria by Biomedica

Size: 1×96 wells
Sensitivity: 0.2 nmol/L
Dynamic Range: 5 – 80 nmol/L
Incubation Time: 4.5 hours
Sample Type: plasma, serum, urine
Sample Size: 20 µL
Alternative Names: Human Endostatin ELISA
For Research Use Only

Controls Included

Conversion Factor: ng/ml to nmol/l:  1 ng/ml = 0.05 nmol/l (MW: 20 kDa)
Specificity: This assay recognizes endogenous and recombinant human Endostatin. There is no cross reactivity with human recombinant Collagen Type XV (COL15).
Cross Reactivity: Human endostatin only. No cross-reactivity with COL15A1.
Reference Values:
Median urine (n=789): 20 pmol/l
Median serum (n=59): 51 pmol/l
Median citrate plasma (n=30): 47 pmol/l
Each laboratory should establish its own reference range for the samples under investigation. Do not change sample type during the study.

Validated for studies of Humans and Rat/Mice.

Assay Principle

The Endostatin ELISA kit is a sandwich enzyme immunoassay for the quantitative determination of Endostatin in human serum, plasma and urine samples. In a first step, standard/control/sample and detection antibody (biotinylated polyclonal rabbit anti-human Endostatin) are pipetted into the wells of the microtiter strips, which are pre-coated with polyclonal goat anti-human Endostatin antibody. Endostatin present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. After a washing step, which removes non-specifically unbound material, the conjugate (streptavidin-HRP) is pipetted into the wells and reacts with the detection antibody. After another washing step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of Endostatin present in the sample. This color change is detectable with a standard microtiter plate reader. The concentration of Endostatin in the sample is determined directly from the dose response curve.

Related Products

Endostatin Mouse/Rat ELISA Assay Kit
Big Endothelin-1 ELISA Assay
Total Soluble Neuropilin-1 ELISA

Learn more about Endostatin and Endostatin Related Studies here:

Endostatin Biomarker Spotlight 

Endostatin Human and Rat/Mice Studies

Additional Information

Assay Background

Endostatin, a 20-kDa C-terminal proteolytic fragment of collagen XVIII, is an endogenous angiogenesis inhibitor localized in the vascular basement membrane in various organs.  The biological functions of the endostatin-network involve SPARC, thrombospondin-1, glycosaminoglycans, collagens, and integrins.   Endostatin is expressed during the progression of renal fibrosis in tubular cells of injured tissue. In renal micro-vascular disease, observed in late stages of patients with chronic kidney disease, increased endostatin levels are possibly the consequence of enhanced extracellular matrix degradation. Thus endostatin may become an important marker for progressive microvascular renal disease in patients with chronic kidney disease. Endostatin levels in blood are also likely to increase in patients with other microvascular tissue injuries, including atherosclerosis, myocardial- and brain ischemia. In ischemic stroke patients, high endostatin plasma levels predict a worse long-term clinical outcome.

Assay Procedure

  • All reagents and samples have to be brought to room temperature (18-26°C) before they can be used in the Endostatin ELISA Assay Kit.
  • Mark position for BLANK/STD/SAMPLE/CTRL (Blank/Standard/Sample/Control) on the protocol sheet.
  • Take microtiter strips out of the aluminium bag, reserve a minimum of one well as blank. Unused strips can be stored with desiccant in the aluminium bag at +4°C (2-8°C) until the expiry date.
  1. Pipette 100 µl ASYBUF (Assay Buffer, natural cap) into each well. Pipette additional 100 µl into well marked as blank.
  2. Add 20 µl of 1+100 diluted STD/SAMPLE/CTRL (Standard/Sample/Control) in duplicate into respective well, except blank.
  3. Add 50 µl AB (Antibody) into each well, swirl gently.
  4. Cover tightly and incubate 3 hours at room temperature (18-26°C).
  5. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer). After final wash, remove remaining WASHBUF by strongly tapping plate against paper towel.
  6. Add 200 µl CONJ (Conjugate, amber cap) into each well.
  7. Cover tightly and incubate for 1 hour at room temperature (18-26°C).
  8. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer). After final wash, remove remaining WASHBUF by strongly tapping plate against paper towel.
  9. Add 200 µl SUB (Substrate, blue cap) into each well.
  10. Incubate for 30 min at room temperature (18-26°C) in the dark.
  11. Add 50 µl STOP (Stop solution, white cap) into each well.
  12. Measure absorbance immediately at 450 nm with reference 630 nm, if available.

Standard Curve

Endostatin ELISA Standard Curve

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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