Thyroperoxidase 蛋白
(2 references)
(2 validations)Quick Overview for Thyroperoxidase 蛋白 (ABIN934717)
抗原
See all Thyroperoxidase (TPO) 蛋白蛋白类型
宿主
资源
纯度
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产品特性
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Purified native Human Thyroid Peroxidase protein
Protein Source: Human thyroid glands
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Thyroid Peroxidase (TPO) (AA 19-846) protein (His tag)
TPO 宿主: 人 宿主: HEK-293 Cells Recombinant > 95 % as determined by Tris-Bis PAGE,> 95 % as determined by HPLC
Thyroid Peroxidase (TPO) (AA 32-834) protein (His tag)TPO 宿主: 小鼠 宿主: HEK-293 Cells Recombinant > 95 % as determined by Tris-Bis PAGE,> 95 % as determined by HPLC
Thyroid Peroxidase (TPO) (AA 672-779) protein (GST tag)TPO 宿主: 人 宿主: 小麦胚 Recombinant WB, ELISA, AA, AP
Thyroid Peroxidase (TPO) (AA 1-933) protein (GST tag)TPO 宿主: 人 宿主: 小麦胚 Recombinant WB, ELISA, AA, AP
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应用备注
- Each Investigator should determine their own optimal working dilution for specific applications.
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限制
- 仅限研究用
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- by
- in.vent Diagnostica GmbH, Hennigsdorf, Germany
- No.
- #102793
- 日期
- 2018.04.17
- 抗原
- 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.
- 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.
生效 #102793 (SDS-PAGE)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Coomassie staining subsequently to SDS-PAGE separation of human TPO1 ABIN934717.]()
Coomassie staining subsequently to SDS-PAGE separation of human TPO1 ABIN934717.
Full Methods -
- by
- in.vent Diagnostica GmbH, Hennigsdorf, Germany
- No.
- #103248
- 日期
- 2018.04.17
- 抗原
- 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.
- 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.
生效 #103248 (Lipid Interaction Assay)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Standard curve and ABIN934717 measurements using LIA.]()
Standard curve and ABIN934717 measurements using LIA.
Full Methods -
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状态
- Lyophilized
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溶解方式
- Reconstitute in distilled water.
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缓冲液
- Lyophilized in 20 mM TRIS, with 50 mM KCl, no preservatives.
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储存液
- Without preservative
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注意事项
- Avoid repeated freeze/thaw cycles.
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储存条件
- -20 °C
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储存方法
- Aliquot and store at -20 °C.
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有效期
- 18 months
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: "A sputum bioassay for airway eosinophilia using an eosinophil peroxidase aptamer." in: Scientific reports, Vol. 12, Issue 1, pp. 22476, (2022) (PubMed).
: "Thyroid peroxidase in human endometrium and placenta: a potential target for anti-TPO antibodies." in: Clinical and experimental medicine, (2020) (PubMed).
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: "A sputum bioassay for airway eosinophilia using an eosinophil peroxidase aptamer." in: Scientific reports, Vol. 12, Issue 1, pp. 22476, (2022) (PubMed).
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- Thyroperoxidase (TPO) (Thyroid Peroxidase (TPO))
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别名
- Thyroid Peroxidase
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背景
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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 -
途径
- Thyroid Hormone Synthesis
抗原
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