Human Pro-Collagen II DuoSet ELISA

Catalog # Availability Size / Price Qty
DY7589-05
Ancillary Products Available
Human Pro Collagen II ELISA Standard Curve
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Product Details
Procedure
Citations (11)
FAQs
Supplemental Products
Reviews (1)

Human Pro-Collagen II DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
For five 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 Pro-Collagen II. The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

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

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

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 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

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)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human Pro Collagen II ELISA Standard Curve

Product Datasheets

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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: Collagen II

Collagens comprise a large family of insoluble extracellular glycoproteins that are essential components of connective tissues such as tendons, ligaments, cartilage, bone and skin. The mature polypeptides are secreted as coiled, left-handed helices that subsequently assemble into rope-like collagen fibers. Collagen I is a fibril-forming collagen that requires N- and C- terminal processing. Collagen IV is a network forming collagen whose C-terminus forms dimers and N-terminus forms tetramers.

Entrez Gene IDs:
1280 (Human); 12824 (Mouse); 25412 (Rat)
Alternate Names:
Alpha-1 type II collagen; ANFH; AOM; cartilage collagen; Chondrocalcin; COL11A3; COL2A1; Collagen 2; collagen alpha-1(II) chain; Collagen II; collagen II, alpha-1 polypeptide; collagen type 2; collagen, type II, alpha 1 (primary osteoarthritis, spondyloepiphysealdysplasia, congenital); collagen, type II, alpha 1; Collagen-2; MGC131516; SEDC

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration 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 plates by adding 300 μL of Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

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, 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 Pro-Collagen II 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.

11 Citations: Showing 1 - 10
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  1. Analgesic, Anti-Inflammatory, and Chondroprotective Activities of Siraitia grosvenorii Residual Extract
    Authors: Lee, YM;Kim, DS;
    International journal of molecular sciences
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Differential Effects of Hypoxia versus Hyperoxia or Physoxia on Phenotype and Energy Metabolism in Human Chondrocytes from Osteoarthritic Compared to Macroscopically Normal Cartilage
    Authors: L Jain, SM Bolam, AP Monk, JT Munro, E Chen, J Tamatea, N Dalbeth, RC Poulsen
    International Journal of Molecular Sciences, 2023-04-19;24(8):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Syringaresinol attenuates osteoarthritis via regulating the NF-kappaB pathway
    Authors: X Wang, D Wang, B Deng, L Yan
    International immunopharmacology, 2023-03-28;118(0):109982.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Formononetin inhibits IL-1beta-induced inflammation in human chondrocytes and slows the progression of osteoarthritis in rat model via the regulation of PTEN/AKT/NF-kappaB pathway
    Authors: C Jia, F Hu, D Lu, H Jin, H Lu, E Xue, D Wu
    International immunopharmacology, 2022-10-25;113(0):109309.
    Species: Human, Rat
    Sample Types: Cell Culture Supernates
  5. Physiologic Mechanical Stress Directly Induces Bone Formation by Activating Glucose Transporter 1 (Glut 1) in Osteoblasts, Inducing Signaling via NAD+-Dependent Deacetylase (Sirtuin 1) and Runt-Related Transcription Factor 2 (Runx2)
    Authors: S Somemura, T Kumai, K Yatabe, C Sasaki, H Fujiya, H Niki, K Yudoh
    International Journal of Molecular Sciences, 2021-08-23;22(16):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Concerted actions by MMPs, ADAMTS and serine proteases during remodeling of the cartilage callus into bone during osseointegration of hip implants
    Authors: J Cassuto, A Folestad, J Göthlin, H Malchau, J Kärrholm
    Bone Rep, 2020-09-11;13(0):100715.
    Species: Human
    Sample Types: Plasma
  7. Nomilin targets the Keap1-Nrf2 signalling and ameliorates the development of osteoarthritis
    Authors: XH Xue, JX Xue, W Hu, FL Shi, Y Yang
    J. Cell. Mol. Med., 2020-06-21;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Regulation of differentiation of annulus fibrosus-derived stem cells using heterogeneous electrospun fibrous scaffolds
    Authors: P Zhou, G Chu, Z Yuan, H Wang, W Zhang, Y Mao, X Zhu, W Chen, H Yang, B Li
    Journal of Orthopaedic Translation, 2020-03-03;26(0):171-180.
    Species: Rabbit
    Sample Types: Cell Culture Supernates
  9. Pharmacological blockade of PCAF ameliorates osteoarthritis development via dual inhibition of TNF-alpha-driven inflammation and ER stress
    Authors: D Chen, D Lu, H Liu, E Xue, Y Zhang, P Shang, X Pan
    EBioMedicine, 2019-11-14;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Knockout of Apolipoprotein E in rabbit promotes premature intervertebral disc degeneration: A new in vivo model for therapeutic approaches of spinal disc disorders
    Authors: A Beierfu beta, H Dietrich, C Kremser, M Hunjadi, A Ritsch, T Rülicke, C Thomé, DS Mern
    PLoS ONE, 2017-11-03;12(11):e0187564.
    Species: Rabbit
    Sample Types: Tissue Homogenates

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Human Pro-Collagen II DuoSet ELISA
By Abby Sukarto on 12/07/2017
Sample Tested: Human tissue