Human Profilin-1 (PFN1) ELISA Kit

Profilin-1 is a ubiquitous actin monomer-binding protein belonging to the profilin family. It is thought to regulate actin polymerization in response to extracellular signals. Deletion of this gene is associated with Miller-Dieker syndrome. Profilin is a ubiquitous 12- to 15-kD protein which inhibits the polymerization of actin. It is thought to function by complexing with unpolymerized actin in vivo. Dissociation of the profilin-actin complex is caused by binding of phosphatidylinositol 4,5-bisphosphate to profilin. Ampe et al. (1988) described the amino acid sequence of human platelet profilin and found it to have 95% homology to the sequence of calf spleen profilin.

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Human Profilin-1 (PFN1) ELISA Kit has high sensitivity and excellent specificity for detection of Human PFN1. No significant cross-reactivity or interference between Human PFN1 and analogues was observed.

This Human Profilin-1 (PFN1) ELISA Kit employs a two-site sandwich ELISA to quantitate PFN1 in samples. An antibody specific for PFN1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPFN1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PFN1 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 PFN1 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.