TSH ELISA Kit

Catalog No. ABIN2641907

Product Details TSH ELISA Kit

Target: TSH (Thyroid Stimulating Hormone (TSH))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.6-24 μIU/mL

Minimum Detection Limit: 0.6 μIU/mL

Application: ELISA

Purpose: For the quantitative determination of rat thyroid stimulating hormone (TSH) concentrations in serum, plasma, tissue homogenates, cell lysates.

Sample Type: Serum, Plasma, Tissue Homogenate, Cell Lysate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of rat TSH.

Cross-Reactivity (Details): Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between the target antigen and all analogues for other species. Therefore, cross reaction may still exist.

Sensitivity: 0.3 μIU/mL

Components:

• Assay plate (12 × 8 coated Microwells)
• Standard (freeze dried)
• Biotin-antibody (100 × concentrate)
• HRP-avidin (100 × concentrate)
• Biotin-antibody Diluent
• HRP-avidin Diluent
• Sample Diluent
• Wash Buffer (25 × concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip (for 96 wells)
• Instruction manual

Material not included:

• Microplate reader capable of measuring absorbance at 450nm, with the correction wavelength set at 540nm or 570nm.
• An incubator which can provide stable incubation conditions up to 37°C ± 0.5°C.
• Squirt bottle, manifold dispenser or automated microplate washer.
• Absorbent paper for blotting the microtiter plate.
• 100mL and 500mL graduated cylinders.
• Deionized or distilled water.
• Pipettes and pipette tips.
• Test tubes for dilution.

Application Details

Application Notes:

• The supplier is only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
• Samples to be used within 5 days may be stored at 2-8°C, otherwise samples must be stored at -20°C (≤ 1 month) or -80°C (≤ 2 months) to avoid loss of bioactivity and contamination.
• Grossly hemolyzed samples are not suitable for use in this assay.
• If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
• Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
• Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts of certain chemicals.
• Owing to the possibility of mismatching between antigens from another resource and antibodies used in this supplier's kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by this supplier's products.
• Influenced by factors including cell viability, cell number and cell sampling time, samples from cell culture supernatant may not be recognized by the kit.
• Fresh samples without long time storage are recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.

Comment:

Detection wavelength: 450 nm

Information on standard material:
Depending on the antigen to be detected, standards can be either native or recombinant protein. The recombinant proteins are being expressed in CHO cells in most cases. Please inquire for more information. The formulation of auxiliary material in the standard is considered proprietary information, however it does not contain any poisonous substance. Proclin 300 (1:3000) is used as preservative.

Information on reagents:
In most cases the stop solution provided is 1 N H2SO4. The formulation of wash solution is proprietary information. None of the components contain (sodium) azide, thimerosal, 2-mercaptoethanol (2-ME) or any other poisonous materials. For the sandwich method kits, the sample diluent, antibody diluent, enzyme diluent and standard all contain BSA.

Information on antibodies:
The antibodies provided in different kits vary in regards to clonality and host. Some antibodies are affinity purified, some are Protein A

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol: This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for TSH has been pre-coated onto a microplate. Standards and samples are pipetted into the wells with a Horseradish Peroxidase (HRP) conjugated antibody specific for TSH. Following a wash to remove any unbound reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TSH bound in the initial step. The color development is stopped and the intensity of the color is measured.

Sample Collection:

• Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4 °C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
• Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2-8 °C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
• Tissue Homogenates: Rinse 100 mg tissue with 1× PBS, homogenize in 1mL of 1× PBS and store overnight at -20 °C. After two freeze-thaw cycles to break the cell membranes, centrifuge the homogenates for 5 minutes at 5000 × g, 2-8 °C. Remove and assay the supernate immediately. Alternatively, aliquot and store samples at -20 °C or -80 °C. Centrifuge the sample again after thawing before the assay. Avoid repeated freeze-thaw cycles.

Calculation of Results:

Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.
Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the target antigen concentration versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

Assay Precision: Intra-assay precision (precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess precision.
Inter-assay precision (precision between assays): Three samples of known concentration were tested in twenty assays to assess precision.

• Intra-assay: CV% less than 8%
• Inter-assay: CV% less than 10%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

Handling Advice:

• The kit should not be used beyond the expiration date on the kit label.
• Do not mix or substitute reagents with those from other lots or sources.
• If samples generate values higher than the highest standard, dilute the samples with Sample Diluent and repeat the assay.
• Any variation in Sample Diluent, operator, pipetting technique, washing technique, incubation time/temperature and kit age can cause variation in binding.
• This assay is designed to eliminate interference by soluble receptors, binding proteins and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.

Storage: 4 °C/-20 °C

Storage Comment: For unopened kit: All the reagents should be kept according to the labels on vials.

Expiry Date: 6 months

References for TSH ELISA Kit (ABIN2641907)

Zhang, Chen, Shao, Xu, Song, Xu, Gao, Hu, Zhao: "A High-Fat Diet Rich in Saturated and Mono-Unsaturated Fatty Acids Induces Disturbance of Thyroid Lipid Profile and Hypothyroxinemia in Male Rats." in: Molecular nutrition & food research, Vol. 62, Issue 6, pp. e1700599, (2018) (PubMed).

Yang, Li, Xu: "Antioxidant therapy improves non-thyroidal illness syndrome in uremic rats." in: Renal failure, Vol. 38, Issue 4, pp. 514-20, (2017) (PubMed).

Rudqvist, Spetz, Schüler, Parris, Langen, Helou, Forssell-Aronsson: "Transcriptional response to 131I exposure of rat thyroid gland." in: PLoS ONE, Vol. 12, Issue 2, pp. e0171797, (2017) (PubMed).

Alrefaie, Awad: "Effect of vitamin D3 on thyroid function and de-iodinase 2 expression in diabetic rats." in: Archives of physiology and biochemistry, Vol. 121, Issue 5, pp. 206-9, (2016) (PubMed).

Yang, Zhang, Liu, Yuan, Song, Shao, Zhou, Yan, Guan, Gao, Zhang, Zhao: "High-Cholesterol Diet Disrupts the Levels of Hormones Derived from Anterior Pituitary Basophilic Cells." in: Journal of neuroendocrinology, Vol. 28, Issue 3, pp. 12369, (2016) (PubMed).

Ashwini, Bobby, Joseph: "Mild hypothyroidism improves glucose tolerance in experimental type 2 diabetes." in: Chemico-biological interactions, Vol. 235, pp. 47-55, (2015) (PubMed).

Ge, Peng, Qi, Chen, Zhou: "Depression-like behavior in subclinical hypothyroidism rat induced by hemi-thyroid electrocauterization." in: Endocrine, Vol. 45, Issue 3, pp. 430-8, (2014) (PubMed).

Hajje, Saliba, Itani, Moubarak, Aftimos, Farès: "Hypothyroidism and its rapid correction alter cardiac remodeling." in: PLoS ONE, Vol. 9, Issue 10, pp. e109753, (2014) (PubMed).

Oltulu, Aktug, Uysal, Turgan, Oktem, Erbas, Yavasoglu, Yavasoglu: "Immunoexpressions of embryonic and nonembryonic stem cell markers (Nanog, Thy-1, c-kit) and cellular connections (connexin 43 and occludin) on testicular tissue in thyrotoxicosis rat model." in: Human & experimental toxicology, (2014) (PubMed).

Jin, Choi, Kim, Pack, Kim, Lee: "Effects of simultaneous combined exposure to CDMA and WCDMA electromagnetic fields on serum hormone levels in rats." in: Journal of radiation research, Vol. 54, Issue 3, pp. 430-7, (2013) (PubMed).

Kim, Paik, Kim, Lee, Lee, Choi, Kim, Pack, Kim, Ahn: "Effects of whole-body exposure to 915 MHz RFID on secretory functions of the thyroid system in rats." in: Bioelectromagnetics, Vol. 34, Issue 7, pp. 521-9, (2013) (PubMed).

Derkach, Moyseyuk, Shpakov: "The influence of prolonged streptozotocin diabetes on the thyroid gland function in rats." in: Doklady. Biochemistry and biophysics, Vol. 451, pp. 217-20, (2013) (PubMed).

Dev, Pal, Pal, Ananthanarayanan, Lalitha, Gaur, Adithan: "Role of ventromedial hypothalamus on energy homeostasis in albino rats: effect of gender." in: Indian journal of physiology and pharmacology, Vol. 56, Issue 2, pp. 107-16, (2013) (PubMed).

Ge, Peng, Hu, Wu: "Impaired learning and memory performance in a subclinical hypothyroidism rat model induced by hemi-thyroid electrocauterisation." in: Journal of neuroendocrinology, Vol. 24, Issue 6, pp. 953-61, (2012) (PubMed).

Spencer, Cowans, Chefetz, Tal, Meiri: "First-trimester maternal serum PP-13, PAPP-A and second-trimester uterine artery Doppler pulsatility index as markers of pre-eclampsia." in: Ultrasound in obstetrics & gynecology : the official journal of the International Society of Ultrasound in Obstetrics and Gynecology, Vol. 29, Issue 2, pp. 128-34, (2007) (PubMed).

Target Details for TSH

Target: TSH (Thyroid Stimulating Hormone (TSH))

Abstract: TSH Products

Synonyms: CG7665 ELISA Kit, DLGR-1 ELISA Kit, DLGR1 ELISA Kit, Dmel\CG7665 ELISA Kit, FSH ELISA Kit, FSH-TSH ELISA Kit, Fsh ELISA Kit, Fsh/CG7665 ELISA Kit, LGR1 ELISA Kit, LH/CG receptor ELISA Kit, RH44949p ELISA Kit, TSH ELISA Kit, dLGR1 ELISA Kit, Leucine-rich repeat-containing G protein-coupled receptor 1 ELISA Kit, Lgr1 ELISA Kit

Target Type: Hormone