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Thyroid Peroxidase (TPO) Protein Protein

TPO Origin: Human Host: Human Native > 95 % pure
Pubmed (1) Independent Validation (2)
Catalog No. ABIN934717
$412.32
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100 μg
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  • Target
    Thyroperoxidase (TPO)
    Protein Type
    Native
    Origin
    • 10
    • 5
    • 2
    Human
    Source
    • 7
    • 5
    • 2
    • 1
    • 1
    • 1
    Human
    Characteristics
    Purified native Human Thyroid Peroxidase protein
    Protein Source: Human thyroid glands
    Purity
    > 95 % pure
  • Application Notes
    Each Investigator should determine their own optimal working dilution for specific applications.
    Restrictions
    For Research Use only
  • Validation #102793 (SDS-PAGE)
    'Independent Validation' Badge
    by
    in.vent Diagnostica GmbH, Hennigsdorf, Germany
    No.
    #102793
    Date
    04/17/2018
    Antigen
    TPO
    Lot Number
    A17091521
    Method validated
    SDS-PAGE
    Positive Control

    Molecular weight (Uniprot P07202: 8 possible isoforms of TPO with a molecular weight between 103 and 67 kDa)

    Negative Control

    none

    Notes

    Passed. Native human TPO protein ABIN934717 appears as a band with a molecular weight of approximately 100kDa after separation on an SDS-PAGE gel.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN934717
    Secondary Antibody
    Full Protocol
    • Dilute 2µg of native human TPO (antibodies-online, ABIN934717, lot A17091521) in 2x sample buffer (126mM Tris-HCl pH6.8, 20% glycerol, 4% SDS, 0.02% bromophenol blue, 0.1M DTT).
    • Boil samples for 5min at 95°C.
    • Separate the sample on a denaturing SERVAGel TG PRiME 12% precast gel (Serva, 43266, V170144) in a Electrophoresis chamber (Hoefer Inc., SE250) with Laemmli buffer for 1h at 250V, 50mA/gel along side 4µl Roti-Mark Tricolor (Carl Roth, 8271, lot 326248051) molecular weight marker.
    • Staining of the separated proteins with InstantBlue Protein Stain (Expedeon, ISB1L, lot 170616330) according to the manufacturer’s recommendations.
    • Image acquisition: 600dpi scan and increasing of color saturation.
    Experimental Notes

    The manufacturer’s information concerning the protein’s purity and molecular weight can be confirmed.

  • Validation #103248 (Lipid Interaction Assay)
    'Independent Validation' Badge
    by
    in.vent Diagnostica GmbH, Hennigsdorf, Germany
    No.
    #103248
    Date
    04/17/2018
    Antigen
    TPO
    Lot Number
    A17091521
    Method validated
    Lipid Interaction Assay
    Positive Control

    Protein concentration (manufacturer's specification)

    Standards and controls for LIA based of purified TPO out of human thyroid

    Negative Control

    Dilution Buffer for LIA

    Deionized water for absorbance at 280nm

    Notes

    Passed. Protein concentration of native human TPO protein ABIN934717 can be confirmed by absorbance at 280nm and by LIA.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN934717
    Secondary Antibody
    Full Protocol
    • Measuring protein concentration using absorbance at 280nm:
      • 100µl sample in a 96-Well UV Microplate (Thermo Fisher Scientific, 8404)
      • Measuring at 280nm
      • Calculation with d of 0.28cm
      • Extinction coefficient assuming all pairs of Cys residues form cysteines: Abs 0.1% (=1g/l)= 1.304
    • Measuring TPO concentration using an in-house coated tube LIA:
      • Sample dilution with assay dilution buffer in 6 steps (10000ng/ml and 100ng/ml as pre-dilution; 50, 25, 12.5, 6.25ng/ml as samples)
      • Pipette 300µl tracer and 200µl standards, controls, or samples per tube in duplicates.
      • Incubation ON at RT.
      • Add 1ml washing solution to each coated tube prior to decanting off the liquid.
      • Add 1 ml washing solution to each coated tube three times and decant off the liquid. Turn the tubes upside down for 5–10min and adsorb remaining liquid with blotting paper.
      • Measurement in a luminometer by automatic injection of LIA reagents (sodium hydrate, azotic acid). Measurement time: 1sec.
    • Calculate the concentration based on the standard curve.
    Experimental Notes
    • The LIA measures human TPO in serum, plasma and other fluids, using one anti-TPO antibody at solid phase and another anti-TPO antibody as tracer. The measurement range is from 10 to 50ng/ml.

    • Absorbance at 280nm was 0.22. The calculated concentration considering the extinction coefficient is 0.6mg/ml (expected was 0.5mg/ml).

    • Regression of TPO LIA standard curve was R2=0.997. Calculated concentration of LIA controls was 28ng/ml (expected 20-30ng/ml) and 14ng/ml (expected 10-15ng/ml).

    • The calculated concentration of the diluted samples was 24ng/ml (expected 25ng/ml) and 13ng/ml (expected 12.5ng/ml). The other dilutions were out of measurement range.

  • Format
    Lyophilized
    Reconstitution
    Reconstitute in distilled water.
    Buffer
    Lyophilized in 20 mM TRIS, with 50 mM KCl, no preservatives.
    Preservative
    Without preservative
    Handling Advice
    Avoid repeated freeze/thaw cycles.
    Storage
    -20 °C
    Storage Comment
    Aliquot and store at -20 °C.
  • Rahnama, Mahmoudi, Kazemnejad, Salehi, Ghahiri, Soltanghoraee, Vafaei, Rezaei, Zarnani: "Thyroid peroxidase in human endometrium and placenta: a potential target for anti-TPO antibodies." in: Clinical and experimental medicine, 2020 (PubMed).

  • Target
    Thyroperoxidase (TPO)
    Synonyms
    MSA, TDH2A, TPX, TPO, msa, tpx, xtpo, LOC100218312, thyroid peroxidase, TPO, Tpo, tpo, LOC100547103
    Background
    Thyroid peroxidase or thyroperoxidase (TPO) is an enzyme expressed mainly in the thyroid that liberates iodine for addition onto tyrosine residues on thyroglobulin for the production of thyroxine (T4) or triiodothyronine (T3), thyroid hormones. In humans, thyroperoxidase is encoded by the TPO gene.
    Description: Human thyroid glands.
    Alternative Names: TPO protein, MSA protein, TPX protein
    Pathways
    Thyroid Hormone Synthesis
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