Overview

  • Product name
    Human BMP-4 ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    HT-29 S/N 5 6.3%
    Inter-assay
    Sample n Mean SD CV%
    HT-29 S/N 3 4.9%
  • Sample type
    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Range
    12 pg/ml - 750 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 89 86% - 91%
    Cell culture extracts 104 97% - 108%
    Tissue Extracts 92 87% - 99%
    Cell culture media 118 110% - 125%
    EDTA Plasma 85 80% - 90%
    Citrate Plasma 87 82% - 93%

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

    BMP-4 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of BMP-4 protein in human serum, plasma, cell culture supernatants, and cell and tissue extracts.


    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.


    BMP-4, also known as Bone morphogenetic protein 4, is a secreted protein of TGF-beta protein superfamily. It forms disulfide-linked homodimer. BMP-4 induces cartilage and bone formation. In general, BMPs are implicated in embryogenesis and morphogenesis of various tissues and organs.


    Sensitivity:


    Samples diluted in Sample Diluent NS = 2.5 pg/ml.


    Samples diluted in 1X Cell Extraction Buffer PTR = 1.5 pg/ml.

  • 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
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Human BMP-4 Capture Antibody 10X 1 x 600µl
    Human BMP-4 Detector Antibody 10X 1 x 600µl
    Human BMP-4 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • Research areas
  • Function
    Induces cartilage and bone formation. Also act in mesoderm induction, tooth development, limb formation and fracture repair. Acts in concert with PTHLH/PTHRP to stimulate ductal outgrowth during embryonic mammary development and to inhibit hair follicle induction.
  • Tissue specificity
    Expressed in the lung and lower levels seen in the kidney. Present also in normal and neoplastic prostate tissues, and prostate cancer cell lines.
  • Involvement in disease
    Defects in BMP4 are the cause of microphthalmia syndromic type 6 (MCOPS6) [MIM:607932]; also known as microphthalmia and pituitary anomalies or microphthalmia with brain and digit developmental anomalies. Microphthalmia is a clinically heterogeneous disorder of eye formation, ranging from small size of a single eye to complete bilateral absence of ocular tissues (anophthalmia). In many cases, microphthalmia/anophthalmia occurs in association with syndromes that include non-ocular abnormalities. MCOPS6 is characterized by microphthalmia/anophthalmia associated with facial, genital, skeletal, neurologic and endocrine anomalies.
    Defects in BMP4 are the cause of non-syndromic orofacial cleft type 11 (OFC11) [MIM:600625]. Non-syndromic orofacial cleft is a common birth defect consisting of cleft lips with or without cleft palate. Cleft lips are associated with cleft palate in two-third of cases. A cleft lip can occur on one or both sides and range in severity from a simple notch in the upper lip to a complete opening in the lip extending into the floor of the nostril and involving the upper gum. OFC11 is an unusual anomaly consisting of a paramedian scar of the upper lip with an appearance suggesting that a typical cleft lip was corrected in utero.
  • Sequence similarities
    Belongs to the TGF-beta family.
  • Cellular localization
    Secreted > extracellular space > extracellular matrix.
  • Information by UniProt
  • Alternative names
    • zgc:100779
    • BMP 2B
    • BMP 4
    • BMP-2B
    • BMP-4
    • BMP2B
    • BMP2B1
    • BMP4
    • BMP4_HUMAN
    • Bone morphogenetic protein 2B
    • Bone morphogenetic protein 4
    • DVR4
    • MCOPS6
    • MGC100779
    • OFC11
    • zbmp-4
    • ZYME
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab231930 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

  • Standard curve comparison between Human BMP-4 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • The BMP-4 standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of BMP-4 were measured in duplicates, interpolated from the BMP-4 standard curves and corrected for sample dilution. Undiluted samples are as follows: SW480 cell culture supernatant 100%, HT-29 cell culture supernatant 50%, HepG2 cell culture supernatant 100%, and naïve culture media (100% RPMI + 10% FBS) 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean BMP-4 concentration was determined to be 535.2 pg/mL in neat SW480 cell culture supernatant, 1035 pg/mL in neat HT-29 cell culture supernatant, 103.3 pg/mL in HepG2 cell culture supernatant, and 24.02 pg/mL in naïve culture media containing 10% FBS.

  • Interpolated concentrations of native BMP-4 in human HT-29 cell extract based on a 150 μg/mL extract load, HepG2 cell extract based on a 300 μg/mL extract load, and colon tissue extract based on a 1,000 μg/mL extract load. The concentrations of BMP-4 were measured in duplicate and interpolated from the BMP-4 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean BMP-4 concentration was determined to be 456.7 pg/mL in HT-29 cell extract, 88.13 pg/mL in HepG2 cell extract, and 22.20 pg/mL in colon tissue extract.

  • The concentrations of BMP-4 were measured in duplicates, interpolated from the BMP-4 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma (citrate) 25%, and plasma (EDTA) 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean BMP-4 concentration was determined to be 193.9 pg/mL in neat serum, 162.0 pg/mL in neat plasma (citrate), and 91.1 pg/mL in neat plasma (EDTA).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean BMP-4 concentration in neat serum was determined to be 163.5 pg/mL with a range of 69.95 – 505.8 pg/mL.

  • The BMP-4 standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.

Protocols

References

ab231930 has not yet been referenced specifically in any publications.

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