| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | PINP |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Human Procollagen I N-terminal peptide (PINP) ELISA Kit has high sensitivity and excellent specificity for detection of Human PINP. No significant cross-reactivity or interference between Human PINP and analogues was observed.
Background
The PINP molecule is similar to PIIINP consisting of three distinct structural domains: Col 1 is on the aminoterminal side of the molecule, while Col 2 and Col 3 are situated on the middle of the helically structured molecule (Kühn et al. 1982). The PINP molecule has a molecular mass of 35 000 and is cleared by scavenger receptors in liver endothelial cells (Melkko et al. 1994). PINP often occurs in circulation in two forms of different molecular sizes. One is identical to the trimeric authentic antigen (intact PINP) whereas the other consists of smaller forms of PINP, resembling a single domain of the proα1(I) chain of PINP and is probably a degradation product of type I procollagen or I pN-collagen. Thus, an assay of intact PINP rather than total PINP appears to be more sensitive in detecting changes in the rate of type I collagen synthesis (Melkko et al. 1996, Risteli & Risteli 1999).
This Procollagen I N-terminal peptide (PINP) 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 Procollagen I N-terminal peptide (PINP) 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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