| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | DKFZp586H0723; K-REV; RAL1B; RAS-related protein RAP1B|Ras family small GTP binding protein RAP1B|small GTP binding protein |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
The deduced 184-amino acid protein is 95% identical to RAP1A. RAP1B shares several properties with the RAS protein, suggesting that it may bind GTP/GDP and have a membrane location.RAP1B is mainly located at the membrane, but translocates to the cytosol upon activation. In activated platelets, RAP1B interacts with the reorganized actin-based cytoskeleton. RAP1B is activated by phosphorylation, increased intracellular Ca(2+), and by agonist-induced stimulation of Gi, which results in the rapid binding of GTP to RAP1B. RAP1B is posttranslationally modified by the geranylgeranylation of cys181. F.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate RAP1B in samples. An antibody specific for RAP1B has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRAP1B present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RAP1B is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of RAP1B bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 5.86 pg/mL |
|---|---|
| Detection Range | 23.44-1500 pg/mL |
| Total Assay Time | 3.5–5 hours |
| Compatible Sample Types | serum, plasma, cell culture supernatant, tissue homogenate |
| Species Reactivity | Human |
| Detection Method | Colorimetric (TMB/HRP) |
| Storage | 4°C (short-term); -20°C (long-term) |
✓ Research-Grade Validation
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with serum, plasma, cell culture supernatant, tissue homogenate. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 5.86 pg/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3.5–5 hours, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant RAP1B standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
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