Mouse CYSTM1 Immunoassay Kit

Mouse CYSTM1 Immunoassay Kit

SKU: 32280

★ Download Datasheet PDF ★

Download MSDS

 

Assay range :   0.156 -10 ng/mL

Size:                 

 

  • INTRODUCTION

    Insulin is a peptide hormone exclusively produced in pancreatic beta-cells. It consists of A chain and B chain, which are linked by two disulfide bridges.
    Insulin is the primary hormone responsible for glucose metabolism. Impaired insulin secretion and insulin resistance are key causes of type 2 diabetes (T2D).

  • PRINCIPLE OF THE ASSAY

    This assay is a quantitative sandwich ELISA. The microplate is pre-coated with a polyclonal antibody specific for mouse CYSTM1. Standards and samples are pipetted into the wells and any mouse CYSTM1 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-labelled polyclonal antibody specific for mouse CYSTM1 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 CYSTM1 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 CYSTM1, 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 assay.

    Insulin (ng/ml)

    Absorbance

    (450 nm)

    Blanked Absorbance

    0

    0.061

    0

    0.025

    0.07

    0.009

    0.075

    0.096

    0.035

    0.2

    0.223

    0.162

    0.5

    0.77

    0.709

    1.5

    2.852

    2.791

     

    B.Sensitivity

    The lowest insulin level that can be measured by this assay is 0.025 ng/ml.

     

    C. Precision

    Intra-assay Precision (Precision within an assay) C.V. < 10%.

    Inter-assay Precision (Precision between assays) C.V. <10%.

     

    D. Recovery

    The recovery of the assay was determined by adding various amounts insulin to a sample. The measured concentration of the spiked sample in the assay was compared to the expected concentration. The average recovery was 92%.

     

    E. Specificity

    Percent of cross reactivity

    Human insulin 100%

    Rat insulin 100%

     

    F. Validation

    Insulin levels were measured at various time points after intra-peritoneal glucose challenge in overnight fasting C57 mice (standard chow fed).

     

©2019 by ImmunoDiagnostics Limited.