Recombinant Cynomolgus Siglec-2/CD22 Fc Chimera Protein, CF

Catalog # Availability Size / Price Qty
10031-SL-050
Recombinant Cynomolgus Siglec-2/CD22 Fc Chimera Protein, CF Bioactivity
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Recombinant Cynomolgus Siglec-2/CD22 Fc Chimera Protein, CF Summary

Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
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 0.07-0.42 μg/mL.
Source
Human embryonic kidney cell, HEK293-derived cynomolgus monkey Siglec-2/CD22 protein
Cynomolgus Monkey Siglec-2/CD22
(Asp20-Arg687)
Accession # EHH59463
IEGRMD Human IgG1
(Pro100-Lys330)
N-terminusC-terminus
Accession #
N-terminal Sequence
Analysis
Asp20
Structure / Form
Disulfide-linked homodimer
Predicted Molecular Mass
102 kDa
SDS-PAGE
115-134 kDa, 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.

10031-SL

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 250 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 °C under sterile conditions after reconstitution.

Data Images

Bioactivity Bioactivity View Larger

Recombinant Cynomolgus Monkey Siglec‑2/CD22 Fc Chimera (Catalog # 10031-SL) supports the adhesion of human red blood cells. The ED50 for this effect is 0.07-0.42 ug/mL.

SDS-PAGE SDS-PAGE View Larger

2 μg/lane of Recombinant Cynomolgus Monkey Siglec‑2/CD22 Fc Chimera was resolved with SDS-PAGE underreducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Bluestaining, showing bands at 115-134 kDa and 230-270 kDa, respectively.

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 sialic acid specific I‑type lectins that are characterized by an extracellular domain (ECD) with an N‑terminal Ig‑like V-type domain followed by varying numbers of Ig‑like C2-type domains (1, 2). 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. In humans, two distinct Siglec-2 cDNAs that arise from differential RNA processing of the same gene have been isolated. The predominant Siglec-2 (Isoform 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 four immunoreceptor tyrosine-based inhibition motifs (ITIMs) (3). The variant Siglec-2 (Isoform CD22-alpha) encodes a 647 aa polypeptide missing two Ig-like C2-type domains and has a truncated (23 aa) cytoplasmic tail (4). Within the ECD, cynomolgus Siglec-2 shares 84%, 55%, and 56% aa sequence identity with human, mouse, and rat Siglec-2, respectively. Siglec-2 is an adhesion molecule that preferentially binds alpha 2,6- linked sialic acid on the same (cis) or adjacent (trans) cells. Interaction of Siglec-2 with trans ligands on opposing cells is found to be favored over the binding of ligands in cis (5). Consistent with a single ligand-binding region, the first two N-terminal Ig-like domains mediated CD22 adhesion with lymphocytes, neutrophils, monocytes, and erythrocytes (6). Besides its role as an adhesion molecule, Siglec-2 is a co-receptor 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 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, 8).

References
  1. Varki, A. and T. Angata (2006) Glycobiology 16:1R.
  2. Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
  3. Wilson, G.L et al. (1991) J. Exp. Med. 173:137.
  4. Stamenkovic, I. and B. Seed (1990) Nature 345:74.
  5. Collins, B.E. et al. (2004) Proc. Natl. Acad. Sci. 101:6104.
  6. Engel, P. et al. (1995) J Exp Med. 181:1581
  7. Ravetch, J.V. and L.L. Lanier (2000) Science 290:84.
  8. Wienands, Y.J. et al. (1999) J. Biol. Chem. 274:18769.
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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