Overview

  • Product name
    Human Pro-Collagen I alpha 1 ELISA Kit, Fluorescent
    See all Procollagen Type 1 kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 8 1.8%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 3%
  • Sample type
    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    3.7 pg/ml
  • Range
    3.91 pg/ml - 16000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 93 91% - 94%
    Cell culture media 99 97% - 101%
    Heparin Plasma 101 94% - 107%
    EDTA Plasma 108 105% - 114%
    Citrate Plasma 106 102% - 110%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Pro-Collagen I alpha 1 in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Pro-Collagen I alpha 1 protein in human serum, plasma, cell culture supernatants, and cell and tissue extract samples.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  CatchPoint Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Type I collagen is the most abundant structural protein of connective tissues such as skin, bone and tendon. It is synthesized as a pro-collagen molecule that is characterized by a 300 nm triple helical domain flanked by globular N- and C-terminal propeptides. Specifically, human Pro-Collagen I alpha 1 consists of a signal peptide (amino acids (aa) 1-22), a propeptide (aa 23-161), the mature chain (aa 162-1218), and another propeptide (aa 1219 – 1464). The non-helical propeptides are removed by procollagen N- and C-proteinase activities so that the mature triple helices can self-assemble into collagen fibrils that provide tensile strength to tissues.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Human Pro-Collagen I alpha 1 Capture Antibody 1 x 600µl
    10X Human Pro-Collagen I alpha 1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Human Pro-Collagen I alpha 1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas
  • Relevance
    Collagens are highly conserved throughout evolution and are characterised by an uninterrupted "Glycine X Y" triplet repeat that is a necessary part of the triple helical structure. Type I collagen (95 kDa) is found in bone, cornea, skin and tendon. Mutations in the encoding gene are associated with osteogenesis imperfecta, Ehlers Danlos syndrome, and idiopathic osteoporosis. Reciprocal translocations between chromosomes 17 and 22, where this gene and the gene for Platelet-derived growth factor beta are located, are associated with a particular type of skin tumor called dermatofibrosarcoma protuberans, resulting from unregulated expression of the growth factor. Procollagen type 1 is mainly synthesized by osteoblasts and, after cleavage of the N- and C-terminal extension peptides, is utilized for collagen fibril deposition in the osteoid tissue.
  • Cellular localization
    Secreted
  • Alternative names
    • Alpha 1 type I collagen
    • Alpha 2 type I collagen
    • COL1A1
    • COL1A2
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229389 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Pro-Collagen I alpha 1 were measured in duplicates, interpolated from the Pro-Collagen I alpha 1 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1%, plasma (citrate) 1%, plasma (EDTA) 1%, and plasma (heparin) 1%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Pro-Collagen I alpha 1 concentration was determined to be 142.1 ng/mL in serum, 135.9 ng/mL in plasma (citrate), 112.1 ng/mL in plasma (EDTA) and 102.1 ng/mL in plasma (heparin).

  • The concentrations of Pro Collagen I alpha 1 were measured in duplicate and interpolated from the Pro Collagen I alpha 1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Pro-Collagen I alpha 1 concentration was determined to be 1.62 ng/mL in IMR-90 extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Pro-Collagen I alpha 1 concentration was determined to be 197.3 ng/mL with a range of 113.0 – 417 ng/mL.

Protocols

References

ab229389 has not yet been referenced specifically in any publications.

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