Human TGF-beta RII | ||||||||||||||||||||||||||||
Human TGF-beta RII DuoSet ELISA Summary
Product Summary This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human TGF-beta RII. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet. Preparation and Storage
Background: TGF-beta RIIMost cell types express three sizes of receptors for TGF-beta. These are designated Type I (53 kDa), Type II (70 - 85 kDa), and Type III (250 - 350 kDa). The Type I receptor is a membrane-bound serine/threonine kinase that apparently requires the presence of the Type II receptor to bind TGF-beta. The Type II receptor is also a membrane-bound serine/threonine kinase that binds TGF-beta1 and TGF-beta 3 with high affinity and TGF-beta 2 with much lower affinity. The Type I and Type II receptors together form a heterodimeric signaling complex that is essential for the transduction of the anti-proliferative signals of TGF-beta. The Type III receptor is a transmembrane proteoglycan with a large extracellular domain and a 43 amino acid residue cytoplasmic domain. The cytoplasmic domain of the Type III receptor lacks an obvious signaling motif and the receptor may not be involved directly in signal transduction.
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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