Human THRSP (Thyroid Hormone Responsive) ELISA Kit (HUES02439)

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HUES02439
€539

Description

ELISA Kit Technical ManualMSDS

Human THRSP (Thyroid Hormone Responsive) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human THRSP . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human THRSP and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human THRSP, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human THRSP. The concentration of Human THRSP in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range0.16-10 ng/mL
Sensitivity0.10 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human THRSP in samples. No significant cross-reactivity or interference between Human THRSP and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(ng/mL)
O.D Average Corrected
102.451
2.481
2.4662.38
51.613
1.641
1.6271.541
2.51.005
0.975
0.990.904
1.250.469
0.501
0.4850.399
0.630.309
0.279
0.2940.208
0.320.201
0.177
0.1890.103
0.160.134
0.144
0.1390.053
00.078
0.094
0.086--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human THRSP were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human THRSP were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.471.114.790.431.174.61
Standard deviation0.030.060.210.030.070.15
C V (%)6.385.414.386.985.983.25

Recovery

The recovery of Human THRSP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)90-10495
EDTA plasma (n=5)95-109101
Cell culture media (n=5)93-10799

Linearity

Samples were spiked with high concentrations of Human THRSP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)93-10993-10697-115
Average (%)10098105
1:4Range (%)89-10179-8988-102
Average (%)948595
1:8Range (%)91-10581-9186-97
Average (%)968692
1:16Range (%)89-10583-9689-101
Average (%)968894

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human THRSP (Thyroid Hormone Responsive) ELISA Kit (HUES02439) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human THRSP (Thyroid Hormone Responsive) ELISA Kit (HUES02439) Protein Information

UniProt Protein Function:THRSP: Plays a role in the regulation of lipogenesis, especially in lactating mammary gland. Important for the biosynthesis of triglycerides with medium-length fatty acid chains. May modulate lipogenesis by interacting with MID1IP1 and preventing its interaction with ACACA. May function as transcriptional coactivator. May modulate the transcription factor activity of THRB. Belongs to the SPOT14 family.
UniProt Protein Details:

Protein type:Unknown function

Chromosomal Location of Human Ortholog: 11q14.1

Cellular Component: nucleoplasm; cytosol

Molecular Function:protein homodimerization activity

Biological Process: transcription, DNA-dependent; regulation of transcription, DNA-dependent; lipid metabolic process; regulation of lipid biosynthetic process

NCBI Summary:The protein encoded by this gene is similar to the gene product of S14, a rat gene whose expression is limited to liver and adipose tissue and is controlled by nutritional and hormonal factors. This gene has been shown to be expressed in liver and adipocytes, particularly in lipomatous modules. It is also found to be expressed in lipogenic breast cancers, which suggests a role in controlling tumor lipid metabolism. [provided by RefSeq, Jul 2008]
UniProt Code:Q92748
NCBI GenInfo Identifier:2842710
NCBI Gene ID:7069
NCBI Accession:Q92748.1
UniProt Secondary Accession:Q92748,B2R4W7,
UniProt Related Accession:Q92748
Molecular Weight:146
NCBI Full Name:Thyroid hormone-inducible hepatic protein
NCBI Synonym Full Names:thyroid hormone responsive
NCBI Official Symbol:THRSP  
NCBI Official Synonym Symbols:S14; Lpgp; THRP; LPGP1; SPOT14  
NCBI Protein Information:thyroid hormone-inducible hepatic protein; SPOT14 homolog; spot 14 protein; lipogenic protein 1; thyroid hormone responsive SPOT14; thyroid hormone responsive (SPOT14 homolog, rat)
UniProt Protein Name:Thyroid hormone-inducible hepatic protein
UniProt Synonym Protein Names:Spot 14 protein; S14; SPOT14
Protein Family:Thyroid hormone-inducible hepatic protein
UniProt Gene Name:THRSP  
UniProt Entry Name:THRSP_HUMAN
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Additional Information

Reactivity:
Human
ELISA Type:
Sandwich
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