| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | SEMA4B; KIAA1745; MGC131831; SEMAC; SemC; sema domain; immunoglobulin domain (Ig); transmembrane domain (TM) and short cytoplasmic domain; 4B; semaphorin 4B |
| 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 Semaphorin-4B (SEMA4B) ELISA Kit has high sensitivity and excellent specificity for detection of Human SEMA4B. No significant cross-reactivity or interference between Human SEMA4B and analogues was observed.
Background
Semaphorin-4B is a protein encoded by the SEMA4B gene. Members of the collapsin/semaphorin gene family have been proposed to act as growth cone guidance signals in vertebrates and invertebrates. To identify candidate molecules involved in axonal pathfinding during mouse embryogenesis, isolated cDNAs encoding five new members of the semaphorin family (Sem A-Sem E). The murine semaphorin genes are differentially expressed in mesoderm and neuroectoderm before and during the time when axons select their pathways in the embryo. In explant cultures, recombinant Sem D/collapsin converts a matrix permissive for axonal growth into one that is inhibitory for neurites of peripheral ganglia. Semaphorins are a diverse family of molecules that may provide local signals to specify territories nonaccessible for growing axons.
This Semaphorin-4B (SEMA4B) 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 Semaphorin-4B (SEMA4B) 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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