Thyroid Peroxidase (TPO) Protein

Details for Product No. ABIN934730, Supplier: Log in to see
Protein Name
  • MSA
  • TDH2A
  • TPX
  • TPO
  • msa
  • tpx
  • xtpo
  • LOC100218312
  • thyroid peroxidase
  • TPO
  • Tpo
  • tpo
  • LOC100547103
Origin
Human
24
14
6
1
Source
Baculovirus infected Insect Cells
22
10
3
2
2
1
1
1
1
Protein Type
Recombinant
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'Independent Validation' Badge
Antigen TPO
Lot Number A13062501
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. Recombinant human TPO protein ABIN934730 appears as a band with a molecular weight of approximately 100kDa after separation on an SDS-PAGE gel.

Primary Antibody ABIN934730
Protocol
  • Dilute 2µg of recombinant TPO (antibodies-online, ABIN934730, lot A13062501) 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 Images
SDS-PAGE validation image for Thyroid Peroxidase (TPO) protein (ABIN934730) Coomassie staining subsequently to SDS-PAGE separation of human TPO1 ABIN934730.
'Independent Validation' Badge
Antigen TPO
Lot Number A13062501
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 recombinant human TPO protein ABIN934730 can be confirmed by absorbance at 280nm and by LIA.

Primary Antibody ABIN934730
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.

Validation Images
Lipid Interaction Assay validation image for Thyroid Peroxidase (TPO) protein (ABIN934730) Standard curve and ABIN934730 measurements using LIA.
Characteristics Purified recombinant Human Thyroid Peroxidase protein
Expression System: Baculovirus in Sf-9 insect cells
Purity > 96 % pure
Protein Name
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.
Alternative Names: TPX protein, MSA protein, TPO protein
Molecular Weight 104 kDa
Pathways Thyroid Hormone Synthesis
Application Notes Each Investigator should determine their own optimal working dilution for specific applications.
Restrictions For Research Use only
Format Liquid
Buffer Supplied as a frozen liquid in 40 mM TRIS HCl buffer, pH 7.6, containing 140 mM NaCl, 0.00 mM Kl and 20 % glycerol.
Handling Advice Avoid repeated freeze/thaw cycles.
Storage -80 °C
Storage Comment Store below -60 °C.
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