Standard Curve
Example of Mouse IgM standard curve in Assay Diluent A1. Plotted are the background-subtracted data.
This standard curve is provided for demonstration only.
Mouse IgM OneStep ELISA Kit
Mouse IgM OneStep ELISA Kit
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Product Details
Product Details
Product Specification
| Antigen | IgM |
| Immunogen | Recombinant Protein |
| Antibody Type | Recombinant mAb |
| Reactivity | Ms |
| Purification | Protein A |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8°C as supplied. |
Kit
| Precision | Intra-assay: 3.9%； Inter-assay: 8.7% |
| Sample type | Serum; Citrate plasma; Cell Culture Supernatant |
| Assay type | Sandwich (quantitative) |
| Sensitivity | 0.302 ng/mL |
| Range | 0.938 ng/mL – 60 ng/mL |
| Recovery | Serum (Balb/c): 104% Citrate plasma (Balb/c): 100% Cell Culture Supernatant: 83% |
| Assay time | 60 minutes |
| Species reactivity | Ms |
Background
Mouse immunoglobulin M (IgM) is a crucial component of the humoral immune system and is the first antibody produced by B-cells in response to an antigenic challenge. It exists in two forms: a pentameric (19S) form found in serum and a monomeric (8S) form on the surface of lymphocytes. The IgM pentamer, which is the predominant form in circulation, consists of five monomeric units and has a molecular weight of approximately 900 kDa, while the hexameric form has a molecular weight of about 1050 kDa. Each monomer of IgM is composed of two heavy chains and two light chains, with the heavy chain having four constant domains (CH1-CH4) and the light chain having one constant domain (CL). The CH4 domain facilitates oligomerization, leading to the formation of penta- or hexamers, and these oligomers are covalently linked via disulfide bonds, resembling a snowflake structure. A joining (J) chain, a 15 kDa polypeptide, is essential for the secretion of IgM into mucosal areas.
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ELISA
Spike Recovery
Recovery of Mouse IgM was assessed by spiking a 60 ng/mL standard into 0.005% serum. Recovery of Mouse IgM was assessed by spiking a 5 ng/mL standard into 0.0067% EDTA-Plasma. Serial dilutions of the spiked sample were prepared, and the percent recovery was calculated for each dilution relative to the expected concentration.
Dilution Linearity
The concentrations of Mouse IgM were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 0.01% concentration. "undiluted" was defined as 0.01% biological sample. The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 627.91 ug/mL in Balb/c Mouse serum.
The mean target concentration was determined to be 336.48 ug/mL in Balb/c Mouse Citrate-plasma.
The mean target concentration was determined to be 579.89 ug/mL in Balb/c Mouse EDTA-plasma.
Dilution Linearity
The concentrations of Mouse IgM were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 0.05% concentration. "undiluted" was defined as 0.05% biological sample. The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 245.38 ug/mL in C57 Mouse serum.
The mean target concentration was determined to be 168.01 ug/mL in C57 Mouse Citrate-plasma.
The mean target concentration was determined to be 175.85 ug/mL in C57 Mouse Heparin-plasma.
Dilution Linearity
The concentrations of Mouse IgM were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 0.05% concentration. "undiluted" was defined as 0.05% biological sample. The subsequent bars represent two-fold serial dilutions of this reference. The mean target concentration was determined to be 260.18 ug/mL in CD-1 Mouse serum.
The mean target concentration was determined to be 190.53 ug/mL in CD-1 Mouse EDTA-plasma.
The mean target concentration was determined to be 96.06 ug/mL in CD-1 Mouse Heparin-plasma.
Intra-Assay Precision (Precision within an assay)
Three samples of known concentration were tested fifteen times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays).
Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed with at least three lots of components.
Determination of Minimum Detectable Dose (MDD)
The MDD was determined using three independent lots of assay components. For each lot, 19 replicate measurements of the diluent (zero calibrator) were performed. The mean (AVERAGE) and standard deviation (STDEV) of the 19 replicates were calculated. The MDD for each lot was then calculated according to the following formula:
MDD = 2 × STDEV + AVERAGE
Cross‑reactivity
Tests for the specificity of various serum and proteins showed no cross-reactivity.
HOOK Effect Threshold
The upper limit of the HOOK effect was established at 100× the highest calibrator concentration (equivalent to a 2-log10 increase). No HOOK effect was observed below this threshold, confirming that samples within this concentration range yield reliable quantitative results without signal depression.
Protocol Diagram
