| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | ACAT2; Acetoacetyl Coenzyme A thiolase; cytosolic acetoacetyl-CoA thiolase |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Gene ID | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
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Features & Benefits
Human Acetyl-CoA acetyltransferase, cytosolic (ACAT2) ELISA Kit has high sensitivity and excellent specificity for detection of Human ACAT2. No significant cross-reactivity or interference between Human ACAT2 and analogues was observed.
Background
ACAT2 is an enzyme involved in lipid metabolism, and it encodes cytosolic acetoacetyl-CoA thiolase. This gene shows complementary overlapping with the 3-prime region of the TCP1 gene in both mouse and human. These genes are encoded on opposite strands of DNA, as well as in opposite transcriptional orientation.As the human TCP1 gene had been assigned to 6q25-q27 by study of somatic cell hybrids and by in situ hybridization, the ACAT2 gene was suspected to be localized to the same chromosome region. Retention of this close linkage during mammalian evolution suggests the possibility of some functional significance. Transcription of both DNA strands at a given locus is common in prokaryotic and viral systems. For examples of overlapping transcriptional units in humans.
This Acetyl-CoA acetyltransferase, cytosolic (ACAT2) 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 Acetyl-CoA acetyltransferase, cytosolic (ACAT2) 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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