Human Siglec-2/CD22 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 Sialic Acid Binding Ig-like Lectin 2 (Siglec-2). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Preparation and Storage

Background: Siglec-2/CD22

Siglec-2 (sialic acid binding Ig-like lectin 2), also known as CD22, is a 130-140 kDa type I transmembrane glycoprotein with seven Ig-like domains in the extracellular region and three ITIM motifs in its cytoplasmic region. Alternate splicing generates an additional isoform that lacks the third and fourth Ig-like domains. Siglec-2 is expressed on B cells, preferentially binds alpha 2-6 linked sialic glycans including CD45 and mediates B cell - B cell interactions. Phosphorylation of Tyr822 within one of the ITIMs is important for Siglec-2 signal transduction through its association with the kinases Lyn, PI 3-Kinase p85 alpha, and SYK.

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.