| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | SEMA4A; RP11-54H19.2; CORD10; FLJ12287; RP35; SEMAB; SEMB; OTTHUMP00000015917; OTTHUMP00000015918; sema domain; immunoglobulin domain (Ig); transmembrane domain (TM) and short cytoplasmic domain; 4A; semaphori |
| 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-4A (SEMA4A) ELISA Kit has high sensitivity and excellent specificity for detection of Human SEMA4A. No significant cross-reactivity or interference between Human SEMA4A and analogues was observed.
Background
SEMA4A is a member of the semaphorin family of soluble and transmembrane proteins. Semaphorins are involved in guidance of axonal migration during neuronal development and in immune responses.Mice treated with Sema4a had enhanced priming of antigen-specific T cells secreting both IL4 and gamma-interferon (IFNG). Treatment of mice with anti-Sema4a early after immunization with myelin oligodendrocyte glycoprotein suppressed the development of experimental autoimmune encephalomyelitis. In these mice, infiltration of mononuclear inflammatory cells was ablated by inhibition of the generation of antigen-specific T cells by anti-Sema4a. Expression cloning determined that the high-affinity receptor for Sema4a is Tim2. Sema4a stimulation of cells expressing Tim2 induced tyrosine phosphorylation of Tim2 but not of Tim3.
This Semaphorin-4A (SEMA4A) 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-4A (SEMA4A) 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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