Mouse SOCS3 (Suppressors Of Cytokine Signaling 3) CLIA Kit (MOES00541)

SKU:
MOES00541
Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
CLIA
€649

Description

ELISA Kit Technical ManualMSDS

Mouse SOCS3 (Suppressors Of Cytokine Signaling 3) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Mouse SOCS3 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse SOCS3 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 SOCS3, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Mouse SOCS3. The concentration of Mouse SOCS3 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection methodChemiluminescence
Detection range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Mouse SOCS3 in samples. No significant cross-reactivity or interference between Mouse SOCS3 and analogues was observed.

Typical Data

As the RLU 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
(pg/mL)
RLU Average Corrected
200055092
56350
5572155693
100021852
26096
2397423946
50011882
10182
1103211004
2504890
5698
52945266
1252640
2576
26082580
62.51324
1298
13111283
31.25667
681
674646
027
29
28--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse SOCS3 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse SOCS3 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)98.36249.10946.3089.10238.88886.88
Standard deviation9.6024.6173.627.2926.5676.54
C V (%)9.769.887.788.1811.128.63

Recovery

The recovery of Mouse SOCS3 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)92-10699
EDTA plasma (n=5)98-114104
Cell culture media (n=5)97-112102

Linearity

Samples were spiked with high concentrations of Mouse SOCS3 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 (%)95-110100-11487-99
Average (%)10310893
1:4Range (%)93-10792-10496-112
Average (%)10099102
1:8Range (%)96-11096-11092-105
Average (%)101103100
1:16Range (%)93-10996-111102-117
Average (%)100101108

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 CLIA 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 Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse SOCS3 (Suppressors Of Cytokine Signaling 3) CLIA Kit (MOES00541) 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Mouse SOCS3 (Suppressors Of Cytokine Signaling 3) CLIA Kit (MOES00541) Protein Information

UniProt Protein Function:SOCS family proteins form part of a classical negative feedback system that regulates cytokine signal transduction. SOCS3 is involved in negative regulation of cytokines that signal through the JAK/STAT pathway. Inhibits cytokine signal transduction by binding to tyrosine kinase receptors including gp130, LIF, erythropoietin, insulin, IL12, GCSF and leptin receptors. Binding to JAK2 inhibits its kinase activity. Suppresses fetal liver erythropoiesis. Regulates onset and maintenance of allergic responses mediated by T-helper type 2 cells. Regulates IL-6 signaling in vivo. Probable substrate-recognition component of a SCF-like ECS (Elongin BC-CUL2/5-SOCS-box protein) E3 ubiquitin-protein ligase complex which mediates the ubiquitination and subsequent proteasomal degradation of target proteins. Seems to recognize IL6ST.
UniProt Code:O35718
NCBI GenInfo Identifier:6671758
NCBI Gene ID:12702
NCBI Accession:NP_031733.1
UniProt Secondary Accession:O35718,P97803, Q3U7X5,
UniProt Related Accession:O35718
Molecular Weight:24,776 Da
NCBI Full Name:suppressor of cytokine signaling 3
NCBI Synonym Full Names:suppressor of cytokine signaling 3
NCBI Official Symbol:Socs3
NCBI Official Synonym Symbols:Cis3; Ef10; Ssi3; Cish3; EF-10; SSI-3
NCBI Protein Information:suppressor of cytokine signaling 3
UniProt Protein Name:Suppressor of cytokine signaling 3
UniProt Synonym Protein Names:Cytokine-inducible SH2 protein 3; CIS-3; Protein EF-10
Protein Family:Suppressor of cytokine signaling
UniProt Gene Name:Socs3
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