Mouse Interleukin-33 (IL-33) ELISA Kit

Mouse Interleukin-33 (IL-33) ELISA Kit

SKU: 32750

★ Download Datasheet PDF 

Download MSDS

 

Assay range: 31.25 - 2000 pg/mL

Kit Size: 96 wells/kit

Other Names: DVS27-related Protein, IL1F11

  • PRINCIPLE OF THE ASSAY

    This assay is a quantitative sandwich ELISA. The microplate is pre-coated with a polyclonal antibody specific for mouse IL-33. Standards and samples are pipetted into the wells and any mouse IL-33 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin labelled polyclonal antibody specific for mouse IL-33 is added to the wells. After wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP substrate solution is added and colour develops in proportion to the amount of mouse IL-33 bound initially. The assay is stopped and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured mouse IL-33, the unknown sample concentration can be interpolated from a reference curve included in each assay.

  • ASSAY PERFORMANCE

    A. Typical representation of standard curve

    The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample assay.  

    Mouse IL-33 (pg/mL)

    Absorbance 

    (450 nm)

    Blanked Absorbance

    0

    0.11

    0

    31.25

    0.145

    0.027

    62.5

    0.179

    0.062

    125

    0.236

    0.118

    250

    0.319

    0.202

    500

    0.65

    0.532

    1000

    1.2

    1.083

    2000

    2.35

    2.233

     

    B. Sensitivity

    The lowest level of mouse IL-33 that can be detected by this assay is 6.5 pg/mL.

     

    C. Precision

    Intra-Assay Precision (Precision within an assay) 

    Three samples of known concentration were tested 8 times on one plate.       

    Sample

    Mean (pg/mL)

    SD (pg/mL)

    CV (%)

    1

    1387.6

    56.4

    4.1

    2

    700.1

    20.7

    6.5

    3

    56.4

    3.0

    5.7

    Intra-Assay Precision (Precision within an assay) 

    Three samples of known concentration were tested 8 times on separate plates.

    Sample

    Mean (pg/mL)

    SD (pg/mL)

    CV (%)

    1

    1425.7

    33.5

    2.4

    2

    713.8

    29.6

    4.2

    3

    146.3

    18.3

    12.5

     

     D. Linearity

    To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse IL-33 were serially diluted with the 1×Assay buffer to produce samples with values within the dynamic range of the assay.

    Serial dilution

    Measured (pg/mL)

    Expected (pg/mL)

    Recovery %

    Neat

    1427.7

    1427.7

    100

    1:2

    703.7

    713.9

    98.6

    1:4

    329.7

    356.9

    92.4

    1:8

    171.2

    178.5

    95.9

     

    F. Validation

    Cell culture supernates:

    Lungs from mice were chopped into 1-2 mm pieces and cultured in 15 mL RPMI supplemented with 10% FBS, 50 μM β-ME, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate or stimulated with 1.0 μg/mL lipopolysaccharide (LPS) for 24 hours. Cell culture medium were removed and assayed for levels of mouse IL-33. In addition, we also collected the cell lysates and measured IL-33 levels and compared with the total protein levels in the cell lysates.       

    Sample

    (cell lysates)

    IL-33 levels/ total protein 

    (pg/mg)

    LPS-stimulated

    381.7

    Unstimulated

    190.9

$450.00Price

For bulk order (QTY: 5 or more), please contact us for quote