Human Ras-related protein Rab-5B (RAB5B) ELISA Kit

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.

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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.

This Human Ras-related protein Rab-5B (RAB5B) ELISA Kit employs a two-site sandwich ELISA to quantitate RAB5B in samples. An antibody specific for RAB5B has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyRAB5B present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RAB5B 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 RAB5B bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.