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Mouse STAT1 (Signal Transducer And Activator Of Transcription 1) ELISA Kit (MOES01474)

SKU:
MOES01474
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P42225
Sensitivity:
0.19ng/mL
Range:
0.31-20ng/mL
ELISA Type:
Sandwich
Reactivity:
Mouse
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Epigenetics and Nuclear Signaling
€649
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Description

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Mouse STAT1 (Signal Transducer And Activator Of Transcription 1) ELISA Kit

The Mouse STAT1 (Signal Transducer and Activator of Transcription 1) ELISA Kit is specifically designed for the accurate measurement of STAT1 levels in mouse serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring dependable and consistent results for a variety of research applications.STAT1 is a vital protein involved in cellular signaling pathways, playing a key role in immune response, inflammation, and cell growth regulation.

Dysregulation of STAT1 has been implicated in various diseases, including cancer, autoimmune disorders, and infectious diseases, making it a valuable biomarker for understanding these conditions and exploring potential treatment options.Overall, the Mouse STAT1 ELISA Kit is a valuable tool for researchers studying the role of STAT1 in health and disease, providing accurate and reliable data for further investigation and discovery.

Assay type:Sandwich
Format:96T
Assay time:4.5h
Reactivity:Mouse
Detection Method:Colormetric
Detection Range:0.31-20 ng/mL
Sensitivity:0.19 ng/mL
Sample Volume Required Per Well:100µL
Sample Type:Serum, plasma and other biological fluids
Specificity:This kit recognizes Mouse STAT1 in samples. No significant cross-reactivity or interference between Mouse STAT1 and analogues was observed.

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 Mouse STAT1. 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 Mouse STAT1 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 Mouse STAT1, 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 Mouse STAT1. The concentration of Mouse STAT1 in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:STAT1: transcription factor of the STAT family. Phosphorylated and activated by receptor-associated kinases downstream of certain receptor tyrosine kinases, GPCRs, and receptors for various interleukins and interferons. Forms homo- or heterodimers that translocate into the nucleus where they regulate transcription. Two alternatively spliced isoforms have been described.
UniProt Protein Details:

Protein type:Transcription factor; DNA-binding

Cellular Component: nucleoplasm; axon; dendrite; nuclear chromatin; cytoplasm; nucleolus; nucleus

Molecular Function:identical protein binding; signal transducer activity; protein binding; enzyme binding; protein homodimerization activity; DNA binding; protein phosphatase 2A binding; sequence-specific DNA binding; double-stranded DNA binding; transcription factor activity; tumor necrosis factor receptor binding; CCR5 chemokine receptor binding

Biological Process: transcription from RNA polymerase II promoter; response to peptide hormone stimulus; response to cAMP; blood circulation; positive regulation of transcription, DNA-dependent; positive regulation of smooth muscle cell proliferation; response to lipopolysaccharide; signal transduction; response to exogenous dsRNA; regulation of transcription, DNA-dependent; tumor necrosis factor-mediated signaling pathway; positive regulation of cell proliferation; negative regulation of viral protein levels in host cell; lipopolysaccharide-mediated signaling pathway; response to nutrient; caspase activation; response to drug; transcription, DNA-dependent; cytokine and chemokine mediated signaling pathway; negative regulation of I-kappaB kinase/NF-kappaB cascade; JAK-STAT cascade; negative regulation of angiogenesis; response to bacterium; cellular response to insulin stimulus; response to mechanical stimulus; response to cytokine stimulus; negative regulation of endothelial cell proliferation; positive regulation of transcription from RNA polymerase II promoter

UniProt Code:P42225
NCBI GenInfo Identifier:1174458
NCBI Gene ID:20846
NCBI Accession:P42225. 1
UniProt Related Accession:P42225
Molecular Weight:32. 2kDa
NCBI Full Name:Signal transducer and activator of transcription 1
NCBI Synonym Full Names:signal transducer and activator of transcription 1
NCBI Official Symbol:Stat1
NCBI Official Synonym Symbols:AA408197; 2010005J02Rik
NCBI Protein Information:signal transducer and activator of transcription 1
UniProt Protein Name:Signal transducer and activator of transcription 1
Protein Family:Signal transducer and activator of transcription
UniProt Gene Name:Stat1
UniProt Entry Name:STAT1_MOUSE

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.DAverageCorrected
202.526
2.582		2.5542.464
101.692
1.74		1.7161.626
51.024
0.988		1.0060.916
2.50.494
0.504		0.4990.409
1.250.298
0.284		0.2910.201
0.630.193
0.191		0.1920.102
0.310.136
0.148		0.1420.052
00.089
0.091		0.09--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse STAT1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse STAT1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)1.062.107.390.992.167.44
Standard deviation0.060.120.340.050.120.33
C V (%)5.665.714.605.055.564.44

Recovery

The recovery of Mouse STAT1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5)85-9791
EDTA plasma (n=5)93-110100
Cell culture media (n=5)88-10495

Linearity

Samples were spiked with high concentrations of Mouse STAT1 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 (%)99-11590-10393-107
Average (%)1069798
1:4Range (%)91-10786-10088-103
Average (%)989394
1:8Range (%)90-10785-9788-101
Average (%)989093
1:16Range (%)90-10482-9486-101
Average (%)988792

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 ×12 strips -20°C, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 µL
Concentrated HRP Conjugate (100×) 1 vial, 120 µL -20°C(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 4°C(shading light)
Stop Solution 1 vial, 10 mL 4°C
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy
  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.

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