| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | RCN2; E6BP; ERC-55; ERC55; TCBP49; reticulocalbin 2; EF-hand calcium binding domain (endoplasmic reticulum calcium-binding protein; 55kD) |
| 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 Reticulocalbin-2 (RCN2) ELISA Kit has high sensitivity and excellent specificity for detection of Human RCN2. No significant cross-reactivity or interference between Human RCN2 and analogues was observed.
Background
Reticulocalbin 2 is a calcium-binding protein located in the lumen of the ER, contains six conserved regions with similarity to a high affinity Ca(+2)-binding motif, the EF-hand. The predicted 317-amino acid RCN2 protein contains an N-terminal signal sequence, 6 copies of the EF-hand motif, and a C-terminal His-Asp-Glu-Leu (HDEL) sequence, which was originally defined as an ER retention motif in yeast proteins. Immunocytolocalization and cell fractionation studies demonstrated that RCN2 is a resident ER protein; the HDEL motif is required for its ER retention. RCN2 was detected by antibodies in all cell lines tested. It binds calcium in vitro. By Western blotting, in vitro translated RCN2 protein has a molecular mass of 55 kD, leading the authors to designate it the 'endoplasmic reticulum calcium-binding protein of 55 kD' (ERC55).
This Reticulocalbin-2 (RCN2) 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 Reticulocalbin-2 (RCN2) 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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