Human Endoglin/CD105 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1097
Ancillary Products Available
Human Endoglin / CD105 ELISA Standard Curve
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Product Details
Procedure
Citations (14)
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Human Endoglin/CD105 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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 Endoglin / CD105. 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

 

Data Example

Human Endoglin / CD105 ELISA Standard Curve

Product Datasheets

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: Endoglin/CD105

Endoglin (CD105) is a transmembrane type III receptor for TGF-beta superfamily ligands and plays an important role in smooth muscle differentiation, angiogenesis, and neovascularization. It is highly expressed on proliferating vascular endothelial cells, chondrocytes, and syncytiotrophoblasts of term placenta. Human Endoglin haploinsufficiency can a cause the vascular disorder, hereditary hemorrhagic telangiectasia type I. Elevated levels of anti-angiogenic soluble Endoglin contribute to pathogenicity in preeclampsia. Endoglin associates with the receptors TGF-beta RII, Activin RIIA or RIIB, BMPR-IA/ALK-3, or BMPR-IB/ALK6 and enhance binding of Activin A, BMP-2, -7, -9, TGF-beta 1, or TGF-beta 3. Endoglin can either enhance or inhibit signaling through these receptor complexes.

Entrez Gene IDs:
2022 (Human); 13805 (Mouse); 497010 (Rat)
Alternate Names:
CD105 antigen; CD105; Endoglin; ENDOsler-Rendu-Weber syndrome 1; ENG; HHT1FLJ41744; ORW; ORW1

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 Endoglin/CD105 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.

14 Citations: Showing 1 - 10
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  1. Sulfasalazine reduces placental secretion of antiangiogenic factors, up-regulates the secretion of placental growth factor and rescues endothelial dysfunction
    Authors: FC Brownfoot, NJ Hannan, P Cannon, V Nguyen, R Hastie, LJ Parry, S Senadheera, L Tuohey, S Tong, TJ Kaitu'u-Li
    EBioMedicine, 2019;0(0):.
    Species: Human
    Sample Types: Tissue Culture Supernates
  2. Angiogenic Properties of Menstrual Stem Cells Are Impaired in Women with a History of Preeclampsia
    Authors: M Varas-Godo, S Acuña-Gall, S Venegas-Du, C Hill, A Caceres-Ve, O Realini, LJ Monteiro, G Zavala, M Khoury, R Romero, G Rice, SE Illanes
    Stem Cells Int, 2019;2019(0):1916542.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Melatonin enhances antioxidant molecules in the placenta, reduces secretion of soluble fms-like tyrosine kinase 1 (sFLT) from primary trophoblast but does not rescue endothelial dysfunction: An evaluation of its potential to treat preeclampsia
    Authors: NJ Hannan, NK Binder, S Beard, TV Nguyen, TJ Kaitu'u-Li, S Tong
    PLoS ONE, 2018;13(4):e0187082.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
    Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
    PLoS ONE, 2015;10(8):e0134619.
    Species: Human
    Sample Types: Plasma
  5. Biomarkers of Host Response Predict Primary End-Point Radiological Pneumonia in Tanzanian Children with Clinical Pneumonia: A Prospective Cohort Study.
    Authors: Erdman L, D'Acremont V, Hayford K, Rajwans N, Kilowoko M, Kyungu E, Hongoa P, Alamo L, Streiner D, Genton B, Kain K
    PLoS ONE, 2015;10(9):e0137592.
    Species: Human
    Sample Types: Plasma
  6. Bone morphogenetic protein-9 suppresses growth of myeloma cells by signaling through ALK2 but is inhibited by endoglin.
    Authors: Olsen O, Wader K, Misund K, Vatsveen T, Ro T, Mylin A, Turesson I, Stordal B, Moen S, Standal T, Waage A, Sundan A, Holien T
    Blood Cancer J, 2014;4(0):e196.
    Species: Human
    Sample Types: Serum
  7. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014;14(0):35.
  8. Inherent phenotypic plasticity facilitates progression of head and neck cancer: endotheliod characteristics enable angiogenesis and invasion.
    Authors: Tong M, Han B, Holpuch A, Pei P, He L, Mallery S
    Exp Cell Res, 2013;319(7):1028-42.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Serum concentrations of soluble Flt-1 are decreased among women with a viable fetus and no symptoms of miscarriage destined for pregnancy loss.
    Authors: Kaitu'u-Lino TJ, Whitehead CL, Ngian GL, Permezel M, Tong S
    PLoS ONE, 2012;7(2):e32509.
    Species: Human
    Sample Types: Serum
  10. Differential expression of Vegfr-2 and its soluble form in preeclampsia.
    Authors: Munaut C, Lorquet S, Pequeux C, Coulon C, Le Goarant J, Chantraine F, Noel A, Goffin F, Tsatsaris V, Subtil D, Foidart JM
    PLoS ONE, 2012;7(3):e33475.
    Species: Human
    Sample Types: Plasma
  11. Performance characteristics of combinations of host biomarkers to identify women with occult placental malaria: a case-control study from Malawi.
    Authors: Conroy AL, Liles WC, Molyneux ME
    PLoS ONE, 2011;6(12):e28540.
    Species: Human
    Sample Types: Plasma
  12. Combinations of Host Biomarkers Predict Mortality among Ugandan Children with Severe Malaria: A Retrospective Case-Control Study.
    Authors: Erdman LK, Dhabangi A, Musoke C, Conroy AL, Hawkes M, Higgins S, Rajwans N, Wolofsky KT, Streiner DL, Liles WC, Cserti-Gazdewich CM, Kain KC
    PLoS ONE, 2011;6(2):e17440.
    Species: Human
    Sample Types: Plasma
  13. Matrix metalloproteinase-14 (MT1-MMP)-mediated endoglin shedding inhibits tumor angiogenesis.
    Authors: Hawinkels LJ, Kuiper P, Wiercinska E
    Cancer Res., 2010;70(10):4141-50.
    Species: Human
    Sample Types: Tissue Homogenates
  14. Endoglin (CD105) as a urinary and serum marker of prostate cancer.
    Authors: Fujita K, Ewing CM, Chan DY, Mangold LA, Partin AW, Isaacs WB, Pavlovich CP
    Int. J. Cancer, 2009;124(3):664-9.
    Species: Human
    Sample Types: Serum

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