Mouse IL-1 alpha CLIA Kit (MOES00033)

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MOES00033
€649

Description

ELISA Kit Technical ManualMSDS

Mouse IL1 alpha (Interleukin 1 Alpha) 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 IL1 alpha . 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 IL1 alpha 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 IL1 alpha, 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 IL1 alpha. The concentration of Mouse IL1 alpha 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 range7.81-500 pg/mL
Sensitivity4.69 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Mouse IL1 alpha CLIA Kit in samples. No significant cross-reactivity or interference between Mouse IL1 alpha CLIA Kit 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
50049655
53419
5153751508
25019658
22338
2099820969
1259841
8951
93969367
62.54381
4643
45124483
31.252347
2251
22992270
15.631279
1221
12501221
7.81726
754
740711
028
30
29--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse IL1 alpha CLIA Kit were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse IL1 alpha CLIA Kit were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)26.5048.55210.7928.1046.53214.55
Standard deviation2.344.9016.103.304.0413.13
C V (%)8.8310.097.6411.748.686.12

Recovery

The recovery of Mouse IL1 alpha CLIA Kit 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)88-10195
EDTA plasma (n=5)92-10699
Cell culture media (n=5)90-10395

Linearity

Samples were spiked with high concentrations of Mouse IL1 alpha CLIA Kit 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 (%)101-11594-10985-99
Average (%)10710192
1:4Range (%)86-10195-10783-97
Average (%)9310290
1:8Range (%)98-11395-10689-104
Average (%)10410096
1:16Range (%)86-9784-9698-114
Average (%)9190104

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 IL1 alpha CLIA Kit (MOES00033) 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 IL1 alpha CLIA Kit (MOES00033) Protein Information

UniProt Protein Function:IL1A: Produced by activated macrophages, IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells. Belongs to the IL-1 family.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Cytokine

Cellular Component: extracellular space; cell surface; plasma membrane; extracellular region; cytosol

Molecular Function:interleukin-1 receptor binding; copper ion binding; cytokine activity

Biological Process: positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of mitosis; positive regulation of interleukin-2 biosynthetic process; cytokine and chemokine mediated signaling pathway; germ cell programmed cell death; positive regulation of JNK cascade; positive regulation of interleukin-6 production; fever; positive regulation of stress-activated MAPK cascade; negative regulation of cell proliferation; positive regulation of angiogenesis; connective tissue replacement during inflammatory response; response to copper ion; positive regulation of cell division; keratinization; positive regulation of prostaglandin secretion; immune response; positive regulation of transcription from RNA polymerase II promoter; regulation of sensory perception of pain; inflammatory response; positive regulation of cytokine secretion

UniProt Code:P01582
NCBI GenInfo Identifier:124298
NCBI Gene ID:16175
NCBI Accession:P01582.1
UniProt Related Accession:P01582
Molecular Weight:31,023 Da
NCBI Full Name:Interleukin-1 alpha
NCBI Synonym Full Names:interleukin 1 alpha
NCBI Official Symbol:Il1a  
NCBI Official Synonym Symbols:Il-1a  
NCBI Protein Information:interleukin-1 alpha; IL-1 alpha
UniProt Protein Name:Interleukin-1 alpha
Protein Family:Interleukin
UniProt Gene Name:Il1a  
UniProt Entry Name:IL1A_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
CLIA
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