| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | TIRAP; FLJ42305; Mal; wyatt; MyD88 adapter-like protein; OTTHUMP00000179130; Toll-interleukin 1 receptor domain-containing adaptor protein; Toll-like receptor adaptor protein; adapter protein wyatt |
| 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
Mouse Toll/Interleukin-1 receptor domain-containing adapter protein (TIRAP) ELISA Kit has high sensitivity and excellent specificity for detection of Mouse TIRAP. No significant cross-reactivity or interference between Mouse TIRAP and analogues was observed.
Background
TIRAP is a TIR adaptor protein involved in the TLR4 signaling pathway of the immune system. It activates NF-kappa-B, MAPK1, MAPK3 and JNK, which then results in cytokine secretion and the inflammatory response. Alternative splicing of this gene results in several transcript variants; however, not all variants have been fully described. MAL, unlike MYD88, does not interact with IRAK1 and is not inhibited by the dominant-negative N-terminal region of IRAK1; however, like MYD88, MAL does, through its TIR domain, interact with and is inhibited in NFKB activation by the dominant-negative form of IRAK2. A dominant-negative form of MAL inhibits TLR4 or lipopolysaccharide activation of NFKB, but not NFKB activation by IL1R1 or IL18R. Immunoprecipitation analysis showed that TLR4 and MAL are constitutively associated.
This Toll/Interleukin-1 receptor domain-containing adapter protein (TIRAP) 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 Toll/Interleukin-1 receptor domain-containing adapter protein (TIRAP) 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 Mouse. 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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