| Field | Specification |
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| Accession Number | |
| Alternative Names | UBE2K; DKFZp564C1216; DKFZp686J24237; E2-25K; HIP2; HYPG; LIG; huntingtin interacting protein 2; ubiquitin carrier protein; ubiquitin-conjugating enzyme E2-25 KDA; ubiquitin-conjugating enzyme E2K; ubiquitin-p |
| 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 Ubiquitin-conjugating enzyme E2 K (UBE2K) ELISA Kit has high sensitivity and excellent specificity for detection of Human UBE2K. No significant cross-reactivity or interference between Human UBE2K and analogues was observed.
Background
HIP2 belongs to the ubiquitin-conjugating enzyme family. It binds selectively to a large region at the N terminus of huntingtin. This interaction is not influenced by the length of the huntingtin polyglutamine tract. This protein has been implicated in the degradation of huntingtin and suppression of apoptosis.
The 200-amino acid HIP2 protein sequence was identical to that of bovine ubiquitin-conjugating enzyme E2-25K. Assays of the interaction of HIP2 with huntingtin revealed that HIP2 binds selectively to a large region at the N terminus of huntingtin. This interaction is not influenced by the length of the huntingtin polyglutamine tract. Western blot analysis detected protein bands of 25, 28, and 45 kD; the 28-kD band appeared to be selective for tissues of the central nervous system.
This Ubiquitin-conjugating enzyme E2 K (UBE2K) 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 Ubiquitin-conjugating enzyme E2 K (UBE2K) 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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