| Field | Specification |
|---|---|
| Alternative Names | T4 |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Rat Thyroxine (T4) ELISA Kit has high sensitivity and excellent specificity for detection of Rat T4. No significant cross-reactivity or interference between Rat T4 and analogues was observed.
Background
T4, the principal thyroid hormone largely bound to transport proteins, especially TBG. Given normal levels of thyroid hormone-binding proteins, hyperthyroidism is characterized by increased levels of circulating T4, hypothyroidism by decreased levels. Exceptions to this parallelism between thyroid status and total T4 concentration are found. Levels of TBG are known to be altered under various physiological, pharmacological and genetic conditions. Elevated T4 levels may be obtained when TBG levels are high, as in pregnancy, acute intermittent porphyria, hyperproteinemia, hereditary TBG elevation and in patients undergoing estrogen therapy or taking oral contraceptives. Total T4 levels may be depressed when TBG levels are low, as in nephrotic, hepatic, gastrointestinal and neoplastic disorders; in acromegaly, hypoproteinemia and hereditary TBG deficiency; and in patients undergoing androgen, testosterone or anabolic steroid therapy.
This Thyroxine (T4) 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 Thyroxine (T4) 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 Rat. 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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