Human Total Survivin | ||||||||||||||||||||||||||||
Human Total Survivin DuoSet IC ELISA Summary
Product Summary This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total Survivin in cell lysates. An immobilized capture antibody specific for Survivin binds both phosphorylated and unphosphorylated Survivin. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format. Preparation and Storage
Background: SurvivinSurvivin is a member of the Inhibitor of Apoptosis (IAP) gene family and contains a single BIR domain. Increased survivin expression at the G2/M checkpoint contributes to tissue homeostasis and differentiation during embryonic and fetal development. Increased expression in differentiated adult tissues, however, frequently results in unchecked cell survival, resulting in cancer. Survivin can protect cells from caspase-induced apoptosis activated in mitosis presumably by binding to Caspase-3, -7, and -9.
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1. Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3. Add 50 μL of Assay Diluent to each well. 4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7. Aspirate and wash 5 times. 8. Add 100 μL Substrate Solution to each well. 9. Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
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