Linearity
Mouse spleen cells were pre-incubated for 24h in the presence of R848 and recombinant IL-2.
Six different cryopreservation mouse spleen cells concentrations, 8 replicates, 1 batch. Data presented the mean spot count, range, and coefficient of variation corresponding to the four cell concentrations.
StepSlim Mouse Specific IgM (HRP) Plus ELISpot Kit
StepSlim Mouse Specific IgM (HRP) Plus ELISpot Kit
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Product Details
Product Details
Product Specification
| Antigen | IgM |
| Antibody Type | Recombinant mAb |
| Reactivity | Mouse |
| Purification | Protein A |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8℃ as supplied. |
Kit
| Precision | Intra-assay: 3.02%； Inter-assay: 3.11% |
| Sample type | PBMCs |
| Assay type | Sandwich (Qualitative) |
| Assay time | 18-24hours |
| Species reactivity | Mouse |
| Plate | Strip |
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.
Picture
Picture
Elispot
Precision
Mouse spleen cells were pre-incubated for 24 h in the presence of R848 and recombinant IL-2.
Following pre-stimulation, the cells were washed and seeded at a density of 5 × 10⁴ cells per well following pre-stimulation, and incubated for 24 hours in a 37°C, 5% CO₂ incubator. Repeat across 8 wells in 3 batches to evaluate inter-batch and intra-batch precision. Data presented include mean spot count, range, and coefficient of variation.
Diagram of spots
After pre-stimulation of mouse spleen cells at 37°C with 5% CO₂ for 24 hours, the cells were incubated with HRP-labelled detection antibodies for 2 hours at room temperature. Then, a colour development substrate was added, followed by image analysis and spot counting.
Protocol Diagram
