Overview

  • Product name

    Human Beta IG-H3 ELISA Kit
    See all TGFBI kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 2.4%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 3.2%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    6.3 pg/ml
  • Range

    18.75 pg/ml - 1200 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 104 103% - 107%
    Serum 99 97% - 101%
    Cell culture extracts 94 92% - 98%
    Cell culture media 88 84% - 90%
    Heparin Plasma 94 91% - 99%
    EDTA Plasma 101 98% - 105%
    Citrate Plasma 100 97% - 104%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Beta IG-H3 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of human beta IG-H3 protein in serum, plasma, cell culture supernatant, and cell extract samples.


    The 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. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:



    • Samples in Sample Diluent NS: 17.59 pg/mL

    • Samples in 1X Cell Extraction Buffer PTR: 6.3 pg/mL

  • Notes

    Human beta IG-H3 is also known as Kerato-epithelin and RGD-CAP (RGD-containing collagen associated protein), is a secreted adhesion molecule that binds to collagens and integrins via covalent and noncovalent interactions. It encodes a 683-amino acid protein containing a secretory signal sequence and four homologous internal domains. Mouse and bovine beta IG-H3 proteins are 90.6% and 92.7% homologous to human beta IG-H3, respectively.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human beta IG-H3 Capture Antibody 1 x 600µl
    10X Human beta IG-H3 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Human beta IG-H3 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Binds to type I, II, and IV collagens. This adhesion protein may play an important role in cell-collagen interactions. In cartilage, may be involved in endochondral bone formation.
  • Tissue specificity

    Highly expressed in the corneal epithelium.
  • Involvement in disease

    Defects in TGFBI are the cause of epithelial basement membrane corneal dystrophy (EBMD) [MIM:121820]; also known as Cogan corneal dystrophy or map-dot-fingerprint type corneal dystrophy. EBMD is a bilateral anterior corneal dystrophy characterized by grayish epithelial fingerprint lines, geographic map-like lines, and dots (or microcysts) on slit-lamp examination. Pathologic studies show abnormal, redundant basement membrane and intraepithelial lacunae filled with cellular debris. Although this disorder usually is not considered to be inherited, families with autosomal dominant inheritance have been identified.
    Defects in TGFBI are the cause of corneal dystrophy Groenouw type 1 (CDGG1) [MIM:121900]; also known as corneal dystrophy granular type. Inheritance is autosomal dominant. Corneal dystrophies show progressive opacification of the cornea leading to severe visual handicap.
    Defects in TGFBI are the cause of corneal dystrophy lattice type 1 (CDL1) [MIM:122200]. Inheritance is autosomal dominant.
    Defects in TGFBI are a cause of corneal dystrophy Thiel-Behnke type (CDTB) [MIM:602082]; also known as corneal dystrophy of Bowman layer type 2 (CDB2).
    Defects in TGFBI are the cause of Reis-Buecklers corneal dystrophy (CDRB) [MIM:608470]; also known as corneal dystrophy of Bowman layer type 1 (CDB1).
    Defects in TGFBI are the cause of lattice corneal dystrophy type 3A (CDL3A) [MIM:608471]. CDL3A clinically resembles to lattice corneal dystrophy type 3, but differs in that its age of onset is 70 to 90 years. It has an autosomal dominant inheritance pattern.
    Defects in TGFBI are the cause of Avellino corneal dystrophy (ACD) [MIM:607541]. ACD could be considered a variant of granular dystrophy with a significant amyloidogenic tendency. Inheritance is autosomal dominant.
  • Sequence similarities

    Contains 1 EMI domain.
    Contains 4 FAS1 domains.
  • Post-translational
    modifications

    Gamma-carboxyglutamate residues are formed by vitamin K dependent carboxylation. These residues are essential for the binding of calcium.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix. May be associated both with microfibrils and with the cell surface.
  • Information by UniProt
  • Alternative names

    • RGD containing collagen associated protein
    • AI181842
    • AI747162
    • Beta ig
    • Beta ig h3
    • Beta ig-h3
    • BGH3_HUMAN
    • Big h3
    • BIGH3
    • CDB1
    • CDG2
    • CDGG1
    • CSD
    • CSD1
    • CSD2
    • CSD3
    • EBMD
    • Kerato epithelin
    • Kerato-epithelin
    • LCD1
    • MGC150270
    • RGD CAP
    • RGD-CAP
    • RGD-containing collagen-associated protein
    • TGFBI
    • TGFBI transforming growth factor, beta induced, 68kDa
    • Transforming growth factor beta induced protein ig h3
    • Transforming growth factor-beta-induced protein ig-h3
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab220651 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

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

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

  • The concentrations of beta IG-H3 were measured in duplicates, interpolated from the beta IG-H3 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:4,000, plasma (citrate) 1:4,000, plasma (EDTA) 1:4,000, and plasma (heparin) 1:4,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean beta IG-H3 concentration was determined to be 1,371.41 ng/mL in neat serum, 1,229.89 ng/mL in neat plasma (citrate), 1,480.27 ng/mL in neat plasma (EDTA), and 1,470.88 ng/mL in neat plasma (heparin).

  • The concentrations of beta IG-H3 were measured in duplicates, interpolated from the beta IG-H3 standard curves and corrected for sample dilution. Undiluted samples are as follows: HUVEC supernatant 8%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean beta IG-H3 concentration was determined to be 18,714.03 pg/mL in neat HUVEC supernatant. HUVEC supernatants were cultured in RPMI base media with 10% fetal bovine serum.

  • The concentrations of beta IG-H3 were measured in duplicate and interpolated from the beta IG-H3 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean beta IG-H3 concentration was determined to be 745.58 pg/mL in U87-MG cell extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean beta IG-H3 concentration was determined to be 1,699.81 ng/mL with a range of 1,014.25 – 3,195.39 ng/mL.

Protocols

References

ab220651 has not yet been referenced specifically in any publications.

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