| Field | Specification |
|---|---|
| Accession Number | |
| Alternative Names | MAD2L2; RP3-330O12.4; MAD2B; REV7; MAD2 (mitotic arrest deficient; yeast; homolog)-like 2; MAD2 homolog; OTTHUMP00000002273; mitotic arrest deficient homolog-like 2 |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Gene ID | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Human Mitotic spindle assembly checkpoint protein MAD2B (MAD2L2) ELISA Kit has high sensitivity and excellent specificity for detection of Human MAD2L2. No significant cross-reactivity or interference between Human MAD2L2 and analogues was observed.
Background
MAD2L2 is a component of the mitotic spindle assembly checkpoint that prevents the onset of anaphase until all chromosomes are properly aligned at the metaphase plate. MAD2L2 is a homolog of MAD2L1. The predicted 211-amino acid MAD2B protein shares 24% and 26% sequence identity with yeast Mad2 and human MAD2L1, respectively, in the conserved regions. RT-PCR analysis revealed that both human MAD2 homologs were expressed at similar high levels in a panel of cell lines. Further binding analyses determined that the interaction of MAD2L2 with ADAM9 is mediated through a proline-rich SH3-ligand domain of ADAM9. Northern blot analysis detected 1.35-kb MAD2L3 transcripts in all tissues tested, with highest expression in testis.
This Mitotic spindle assembly checkpoint protein MAD2B (MAD2L2) ELISA kit is validated for use with Cell culture supernatants, Serum, Plasma, Other biological fluids. Samples should be collected, processed, and stored correctly to preserve analyte integrity — avoid repeated freeze-thaw cycles and centrifuge to remove particulates before use. Dilute samples exceeding the kit's detection range using the supplied assay diluent. Hemolytic, icteric, or lipemic samples may affect assay performance and should be tested with caution.
This is a sandwich ELISA kit employing an HRP (horseradish peroxidase)-conjugated secondary antibody paired with a TMB (3,3′,5,5′-tetramethylbenzidine) colorimetric substrate. In the sandwich format, the target analyte Mitotic spindle assembly checkpoint protein MAD2B (MAD2L2) captured on the microplate surface is detected by the conjugated antibody, generating a colorimetric signal proportional to analyte concentration. The reaction is stopped and absorbance measured at 450 nm on a standard microplate reader.
The complete protocol, from sample addition to final plate reading, requires approximately 3–5 hours. This includes two incubation periods (analyte binding and detection antibody steps), intermediate wash cycles to remove unbound material, 15–30 minutes of TMB substrate development, and final stop-solution addition before absorbance reading. Exact timing will vary with experience level and the number of samples processed in parallel.
Required equipment: (1) a microplate spectrophotometer capable of reading absorbance at 450 nm (reference wavelength 540–570 nm recommended for background correction); (2) precision single-channel or multichannel pipettes; (3) a plate washer or multichannel aspirator; (4) a microcentrifuge for sample clarification; and (5) a 37°C incubator or stable room-temperature environment. No fluorescence or luminescence reader is required — standard colorimetric plate readers are fully compatible with this kit.
This ELISA kit is formally validated for Human. Cross-reactivity with species not listed in the specification has not been independently characterized. Variability in protein sequence homology across species means that performance in unlisted species cannot be guaranteed without additional validation. For cross-species detection requirements or non-standard sample matrices, please contact BioHippo support or refer to the manufacturer's technical team for guidance.
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