| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | RAP1B; DKFZp586H0723; K-REV; RAL1B; RAS-related protein RAP1B; Ras family small GTP binding protein RAP1B; small GTP binding protein |
| 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 Ras-related protein Rap-1b (RAP1B) ELISA Kit has high sensitivity and excellent specificity for detection of Human RAP1B. No significant cross-reactivity or interference between Human RAP1B and analogues was observed.
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. Further modifications cause proteolytic removal of the last 3 C-terminal amino acids, followed by partial methylation of the remaining terminal cysteinyl carboxyl group.
This Ras-related protein Rap-1b (RAP1B) 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 Ras-related protein Rap-1b (RAP1B) 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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