| Field | Specification |
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| Mfr No | |
| Accession Number | |
| Alternative Names | RAB5B |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Gene ID | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
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Features & Benefits
Human Ras-related protein Rab-5B (RAB5B) ELISA Kit has high sensitivity and excellent specificity for detection of Human RAB5B. No significant cross-reactivity or interference between Human RAB5B and analogues was observed.
Background
A subset of these GTPases plays a central role in membrane trafficking. Each of the several proteins of this subfamily that have been identified is thought to regulate vesicular trafficking at a specific subcellular compartment. The subcellular location of several RAB proteins has been determined by immunohistochemical methods. For example, RAB2 is found in the intermediate recycling pathway between the endoplasmic reticulum and the Golgi complex. RAB6 is distributed in the medial and trans Golgi. RAB4 and RAB5A are associated with the plasma membrane and early endosomes. The newly identified RAB protein was 81% identical to human RAB5, the canine counterpart of which had been localized to the plasma membrane and early endosomes. RAB5B is presumably involved in vesicular trafficking at the plasma membrane.
This Ras-related protein Rab-5B (RAB5B) 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 Rab-5B (RAB5B) 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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