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Human Endostatin DuoSet ELISA

Catalog #: DY1098
16 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY1098
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human Endostatin ELISA Standard Curve
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Product Details
Procedure
Citations (16)
FAQs
Supplemental Products
Reviews (1)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human Endostatin DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Endostatin. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet ELISA.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

 

Scientific Data

N/A Human Endostatin ELISA Standard Curve View Larger

Human Endostatin ELISA Standard Curve

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Endostatin

Endostatin is a 20 kDa proteolytic fragment of the C-terminal, non-collagenous (NC1) domain of type XVIII Collagen. It was originally identified as a factor produced by murine hemangioendothelioma cells that could specifically inhibit endothelial cell proliferation and angiogenesis. Although the molecular signals that trigger the release of Endostatin from type XVIII Collagen are not well understood, multiple proteases have been suggested to be involved in its generation including Cathepsins S, B, L, and V, Elastase, and matrix metalloproteinases (MMPs)-2, -7, and -9. Endostatin is of particular interest as it has been shown to inhibit the growth of many primary and metastatic tumors. It may also be involved in down-regulating angiogenesis during physiological processes such as wound healing and the establishment of placental circulation. The anti-angiogenic activity of Endostatin is attributable to its ability to inhibit endothelial cell proliferation and suppress VEGF-and FGF basic-induced endothelial cell migration and adhesion. Many of these effects are thought to be mediated by interactions between Endostatin and endothelial cell-expressed Transglutaminase 2, Heparin, and Integrins alpha 5 beta 1 and alpha V beta 3.

Entrez Gene IDs:
80781 (Human); 12822 (Mouse)
Alternate Names:
antiangiogenic agent; COL18A1; collagen alpha-1(XVIII) chain; collagen, type XVIII, alpha 1; Endostatin; FLJ27325; FLJ34914; human type XVIII collagen10endostatin; KNO; KNO1MGC74745; Knobloch syndrome, type 1; KS; multi-functional protein MFP

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) in PBS without carrier protein. Immediately coat a 96-well microplate with 100 µL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 µL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

    Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
     
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

Assay Procedure

  1. Add 100 µL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 µL of the Detection Antibody, diluted in Reagent Diluent (as recommended in the product datasheet), to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 µL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 µL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Endostatin DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

16 Citations: Showing 1 - 10
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  1. Effects of an Intervention with Selenium and Coenzyme Q10 on Five Selected Age-Related Biomarkers in Elderly Swedes Low in Selenium: Results That Point to an Anti-Ageing Effect-A Sub-Analysis of a Previous Prospective Double-Blind Placebo-Controlled Randomised Clinical Trial
    Authors: Alehagen, U;Alexander, J;Aaseth, JO;Larsson, A;Svensson, E;Opstad, TB;
    Cells
    Species: Human
    Sample Types: Plasma
  2. In Vitro Comparison of Lymphangiogenic Potential of Hypoxia Preconditioned Serum (HPS) and Platelet-Rich Plasma (PRP)
    Authors: J Jiang, X Cong, S Alageel, U Dornseifer, AF Schilling, E Hadjipanay, HG Machens, P Moog
    International Journal of Molecular Sciences, 2023-01-19;24(3):.
    Species: Human
    Sample Types: Hypoxia Preconditioned Serum
  3. Prediction of emergency cerclage outcomes in women with cervical insufficiency: The role of inflammatory, angiogenic, and extracellular matrix-related proteins in amniotic fluid
    Authors: KN Lee, KH Park, YM Kim, I Cho, TE Kim
    PLoS ONE, 2022-05-10;17(5):e0268291.
    Species: Human
    Sample Types: Amniotic Fluid
  4. Antibody microarray analysis of amniotic fluid proteomes in women with cervical insufficiency and short cervix, and their association with pregnancy latency length
    Authors: S Hong, KH Park, YE Lee, JE Lee, YM Kim, E Joo, I Cho
    PLoS ONE, 2022-02-07;17(2):e0263586.
    Species: Human
    Sample Types: Amniotic Fluid
  5. A Comparison between Endostatin and Conventional Biomarkers on 30-Day Mortality and Renal Replacement Therapy in Unselected Intensive Care Patients
    Authors: T Ruge, A Larsson, M Lipcsey, J Tydén, J Johansson, M Eriksson
    Biomedicines, 2021-11-03;9(11):.
    Species: Human
    Sample Types: Plasma
  6. Biomarkers of Pulmonary Hypertension Are Altered in Children With Down Syndrome and Pulmonary Hypertension
    Authors: M Griffiths, J Yang, D Vaidya, M Nies, S Brandal, DD Ivy, F Hickey, K Wolter-War, ED Austin, M Mullen, MW Pauciulo, KA Lutz, EB Rosenzweig, R Hirsch, D Yung, WC Nichols, AD Everett
    The Journal of pediatrics, 2021-10-21;0(0):.
    Species: Human
    Sample Types: Plasma
  7. Exploring the potential effect of paricalcitol on markers of inflammation in de novo renal transplant recipients
    Authors: HK Pihlstrøm, T Ueland, AE Michelsen, P Aukrust, F Gatti, C Hammarströ, M Kasprzycka, J Wang, G Haraldsen, G Mjøen, DO Dahle, K Midtvedt, IA Eide, A Hartmann, H Holdaas
    PLoS ONE, 2020-12-16;15(12):e0243759.
    Species: Human
    Sample Types: Plasma
  8. The role of EMMPRIN/CD147 in regulating angiogenesis in patients with psoriatic arthritis
    Authors: MA Rahat, M Safieh, E Simanovich, E Pasand, T Gazitt, A Haddad, M Elias, D Zisman
    Arthritis Res Ther, 2020-10-14;22(1):240.
    Species: Human
    Sample Types: Serum
  9. Plasma biomarkers of the amyloid pathway are associated with geographic atrophy secondary to age-related macular degeneration
    Authors: K Lashkari, GC Teague, U Beattie, J Betts, S Kumar, MM McLaughlin, FJ López
    PLoS ONE, 2020-08-07;15(8):e0236283.
    Species: Human
    Sample Types: Plasma
  10. Immune and Inflammatory Proteins in Cord Blood as Predictive Biomarkers of Retinopathy of Prematurity in Preterm Infants
    Authors: YJ Park, SJ Woo, YM Kim, S Hong, YE Lee, KH Park
    Invest. Ophthalmol. Vis. Sci., 2019-09-03;60(12):3813-3820.
    Species: Human
    Sample Types: Plasma
  11. Blood inflammatory and endothelial markers in women with von Willebrand disease
    Authors: I Govorov, K Bremme, A Larsson, M Holmström, E Komlichenk, R Chaireti, M Mints
    PLoS ONE, 2019-01-10;14(1):e0210544.
    Species: Human
    Sample Types: Plasma
  12. Serum Biomarkers of Myocardial Remodeling and Coronary Dysfunction in Early Stages of Hypertrophic Cardiomyopathy in the Young
    Authors: E Fernlund, T Gyllenhamm, R Jablonowsk, M Carlsson, A Larsson, J Ärnlöv, P Liuba
    Pediatr Cardiol, 2017-03-30;0(0):.
    Species: Human
    Sample Types: Serum
  13. Distinct platelet packaging, release, and surface expression of proangiogenic and antiangiogenic factors on different platelet stimuli.
    Authors: Chatterjee M, Huang Z, Zhang W, Jiang L, Hultenby K, Zhu L, Hu H, Nilsson GP, Li N
    Blood, 2011-02-17;117(14):3907-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Normal ranges of angiogenesis regulatory proteins in human platelets.
    Authors: Peterson JE, Zurakowski D, Italiano JE
    Am. J. Hematol., 2010-07-01;85(7):487-93.
    Species: Human
    Sample Types: Cell Lysates
  15. Angiogenesis and lymphangiogenesis are downregulated in primary breast cancer.
    Authors: Boneberg EM, Legler DF, Hoefer MM, Ohlschlegel C, Steininger H, Fuzesi L, Beer GM, Dupont-Lampert V, Otto F, Senn HJ, Furstenberger G
    Br. J. Cancer, 2009-08-18;101(4):605-14.
    Species: Human
    Sample Types: Serum
  16. Pretreatment serum endostatin as a prognostic indicator in metastatic gastric carcinoma.
    Authors: Woo IS, Kim KA, Jeon HM, Hong SH, Rho SY, Koh SJ, Lee MA, Byun JH, Kang JH, Hong YS, Lee KS, Cho CS, Choi MG, Chung IS
    Int. J. Cancer, 2006-12-15;119(12):2901-6.
    Species: Human
    Sample Types: Serum

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Human Endostatin DuoSet ELISA
By satyendra tripathi on 06/23/2018
Sample Tested: EDTA Plasma