Rat Thyroid Stimulating Hormone,TSH ELISA Kit

Instructions
Code CSB-E05115r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This ELISA kit is used for quantitative detection of rat thyroid-stimulating hormone (TSH), establishing normal detection range, validation of antibody array results. TSH is secreted by the pituitary gland and stimulates thyroid gland growth and its synthesis and secretion of thyroid hormones. It is critical for normal development and metabolism. TSH levels are negatively regulated by thyroid hormone (T3 and T4). TSH is regarded as the primary biomarker for evaluating thyroid function and can help to guide treatment with levothyroxine for patients with hypothyroidism.

Based on the Sandwich-ELISA principle and enzyme-substrate chromogenic reaction property, the users can measure the content of rat TSH in serum, plasma, tissue homogenates, or cell lysates. This rat TSH ELISA kit is quality tested with high specificity, excellent sensitivity, precision less than 15%, good linearity, and high recovery. Click the product instructions for more validation information.

Target Name Thyroid Stimulating Hormone,TSH
Alternative Names TshbThyrotropin subunit beta ELISA kit; Thyroid-stimulating hormone subunit beta ELISA kit; TSH-B ELISA kit; TSH-beta ELISA kit; Thyrotropin beta chain ELISA kit
Abbreviation TSH
Uniprot No. P04652
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, tissue homogenates, cell lysates
Detection Range 0.6 μIU/mL-24 μIU/mL
Sensitivity 0.3 μIU/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Signal Transduction
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<15%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<15%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of rat TSH in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 93  
Range % 87-98  
1:2 Average % 87  
Range % 84-90  
1:4 Average % 91  
Range % 85-96  
1:8 Average % 95  
Range % 90-99  
Recovery
The recovery of rat TSH spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 96 91-100  
EDTA plasma (n=4) 93 88-99  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
μIU/ml OD1 OD2 Average    
24 2.756 2.781 2.769    
12 1.997 1.988 1.993    
6 1.142 1.124 1.133    
3 0.634 0.613 0.624    
1.2 0.244 0.253 0.249    
0.6 0.115 0.117 0.116    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-rat TSH antibody.
  • Standard 7 x 0.5 ml--- Dilute the standard at dilution series, read the OD values, and then draw a standard curve.
  • HRP-conjugated TSH antibody 1 x 6 ml --- Bind to the TSH in the sample or in the standard and react with the TMB substrate to make the solution chromogenic.
  • Wash Buffer (20 x concentrate) 1 x 15 ml --- Wash away unbound or free substances.
  • Substrate A1 x 7 ml --- Mix with Substrate B, and the mixture reacts with HRP to make the solution appear blue.
  • Substrate B 1 x 7 ml --- Mix with Substrate A, and the mixture reacts with HRP to make the solution appear blue.
  • Stop Solution 1 x 7 ml --- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 600 nm - 630 nm.
  • An incubator that can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

Customer Reviews and Q&A

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 Q&A
Q:

Please let us know delivery time and the sulfuric acid content of [stop solution] .

A:
Thanks for your inquiry. The sulfuric acid content of [stop solution] is 4N. Delivery time is usually around 3-5 business days.

Target Background

Function
(From Uniprot)
Indispensable for the control of thyroid structure and metabolism.
Gene References into Functions
  1. These data add new insight to an important role of T3 in rapidly regulating TSH beta gene expression at posttranscriptional level. PMID: 21078364
  2. TSH action stimulates the phosphorylation of hsp-90 in FRTL-5 thyroid cells PMID: 16910874
  3. Results clearly show that there are diurnal changes in TSH expression and accumulation in rat PT-TSH cells and suggest that these fluctuations are regulated by melatonin. PMID: 17927662
Subcellular Location Secreted
Protein Families Glycoprotein hormones subunit beta family
Database Links

KEGG: rno:25653

STRING: 10116.ENSRNOP00000022575

UniGene: Rn.9966

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