Recombinant Human Siglec-2/CD22 His-tag Avi-tag Protein, CF

Biotinylated
Catalog # Availability Size / Price Qty
AVI10191-050
Recombinant Human Siglec-2/CD22 His-tag Avi-tag Protein, CF Binding Activity
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Recombinant Human Siglec-2/CD22 His-tag Avi-tag Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA. When Human Siglec-2/CD22 (Epratuzumab, Novus Catalog # NBP2-75189) is immobilized at 0.5 µg/mL (100 µL/well), Biotinylated Recombinant Human Siglec-2/CD22 His-tag Avi-tag (Catalog # AVI10191) binds with an ED50 of 0.3-2.4 ng/mL. Measured by the ability of the immobilized protein to support the adhesion of human red blood cells. Kelm, S. et al. (1994) Current Biology 4:965. The ED50 for this effect is 0.1-0.8 μg/mL.
Source
Chinese Hamster Ovary cell line, CHO-derived human Siglec-2/CD22 protein
Human Siglec-2/CD22
(Asp20-Arg687)
Accession # CAA42006.1
GGGSGGGSGGGSHHHHHHHAvi-tag
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
79 kDa
Structure / Form
Biotinylated via Avi-tag
Predicted Molecular Mass

79 kDa

SDS-PAGE
92-118 kDa, under reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AVI10191

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Data Images

Binding Activity Binding Activity View Larger

When Human Siglec-2/CD22 (Epratuzumab, Novus Catalog # NBP2-75189) is immobilized at 0.5 μg/mL, 100 μL/well, Biotinylated Recombinant Human Siglec-2/CD22 His-tag Avi-tag (Catalog # AVI10191) binds with an ED50 of 0.3-2.4 ng/mL.

SDS-PAGE SDS-PAGE View Larger

2 μg/lane of Biotinylated Recombinant Human Siglec-2/CD22 His-tag Avi-tag (AVI10191) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 92-118 kDa.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Siglec-2/CD22

Siglecs are type I transmembrane proteins that belong to the immunoglobulin (Ig) superfamily and function as mammalian lectins (1). They are characterized by an extracellular domain consisting of various numbers of Ig domains with a conserved N-terminal V-set Ig ligand-binding domain. This binds species-specific sialic acid motifs on protein and lipid scaffolds to regulate intracellular signaling pathways (2). The cytoplasmic tail has signaling motifs, in most cases immunoreceptor tyrosine-based inhibitory motif (ITIM) (3). Human Siglec-2, also known as B cell antigen CD22 or B lymphocyte cell adhesion molecule (BL-CAM), is a B cell restricted glycoprotein that is expressed in the cytoplasm of progenitor B and pre B cells and on the surface of mature B cells. Two distinct human Siglec-2/CD22 cDNAs that arise from differential RNA processing of the same gene have been isolated. The predominant Siglec-2/CD22 beta encodes an 847 amino acid (aa) polypeptide with a hydrophobic signal peptide, an N-terminal Ig-like V type domain, six Ig-like C2 type domains, a transmembrane region and a cytoplasmic tail with 4 immunoreceptor tyrosine-based inhibition motifs (ITIMs) (4). The variant Siglec-2/CD22 alpha encodes a 647 aa polypeptide missing two Ig-like C2 type domains and has a truncated (23 aa) cytoplasmic tail (5). Mature human Siglec-2 beta consists of a 668 amino acid (aa) extracellular domain (ECD), a 19 aa transmembrane segment, and a 141 aa cytoplasmic domain. Within the ECD, human Siglec-2 shares 59% and 58% aa sequence identity with the mouse and rat Siglec-2, respectively. Siglec-2/CD22 is an adhesion molecule that preferentially binds alpha 2,6- linked sialic acid on the same (cis) or adjacent (trans) cells. Interaction of CD22 with trans ligands on opposing cells was found to be favored over the binding of ligands in cis (6). Besides its role as an adhesion molecule, Siglec-2/CD22 is a coreceptor that physically interacts with B-cell receptor (BCR) and is rapidly phosphorylated upon BCR ligation. It negatively regulates BCR signals by recruiting tyrosine phosphatase SHP-1 to its ITIMs. Phosphorylated Siglec-2/CD22 can also interact with other intracellular effector proteins such as Syk, PLC gamma, PI3 kinase and Grb-2, suggesting it may play a role in positive signaling (7-9). Another function of CD22 is that it mediates the anti-phagocytic effect of alpha 2,6-linked sialic acid, and inhibition of CD22 promotes the clearance of myelin debris, amyloid-beta oligomers and alpha -synuclein fibrils in vivo(10). CD22 also plays a role in autoimmunity and has great potential for CD22-based immunotherapeutics for the treatment of autoimmune diseases such as systemic lupus erythematosus (SLE) (11).

References
  1. Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
  2. Poe, J.C. and T.F. Tedder (2012) Trends Immunol. 33:413.
  3. Meyer, S.J. et al. (2018) Front. Immunol. 9:2820.
  4. Wilson, G.L et al. (1991) J. Exp. Med. 173:137.
  5. Stamenkovic, I. and B. Seed (1990) Nature 345:74.
  6. Collins, B.E. et al. (2004) Proc. Natl. Acad. Sci. 101:6104.
  7. Crocker, P.R. and A. Varki (2001) Immunology 103:137.
  8. Ravetch, J.V. and L.L. Lanier (2000) Science 290:84.
  9. Wienands, Y.J. et al. (1999) J. Biol. Chem. 274:18769.
  10. Pluvinage, J.V. et al. (2019) Nature. 5568:7751.
  11. Clark, E.A. et al. (2018) Front. Immunol. 9:2235.
Long Name
Sialic Acid Binding Ig-like Lectin 2
Entrez Gene IDs
933 (Human); 12483 (Mouse); 102130968 (Cynomolgus Monkey)
Alternate Names
B-cell receptor CD22; BL-CAM; B-lymphocyte cell adhesion molecule; CD22 antigenMGC130020; CD22 molecule; CD22; sialic acid binding Ig-like lectin 2; Sialic acid-binding Ig-like lectin 2; Siglec2; Siglec-2; SIGLEC2FLJ22814; T-cell surface antigen Leu-14

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