Human Siglec‑2 LlamabodyTM VHH His-tag Alexa Fluor® 594‑conjugated Antibody

Catalog #: LFAB10732T Datasheet / COA / SDS
Recombinant Monoclonal Llama VHH domain
Catalog # Availability Size / Price Qty
LFAB10732T-100UG
Detection of Siglec-2/CD22 in HEK293 Human Cell Line Transfected with Human Siglec-2/CD22 and eGFP by Flow Cytometry.
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Human Siglec‑2 LlamabodyTM VHH His-tag Alexa Fluor® 594‑conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects human Siglec-2/CD22 in direct ELISAs.
Source
Recombinant Monoclonal Llama VHH domain Clone # L007.2.5N
Purification
His-tag purified from cell culture supernatant
Immunogen
Chinese Hamster Ovary cell line CHO-derived human Siglec-2/CD22
Asp20-Arg687
Accession # CAA42006.1
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 594 (Excitation= 590 nm, Emission= 617 nm)

Applications

Recommended Concentration
Sample
Flow Cytometry
0.25-1 µg/106 cells
HEK293 human embryonic kidney cell line transfected with human Siglec-2/CD22 and eGFP

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry View Larger

Detection of Siglec-2/CD22 in HEK293 Human Cell Line Transfected with Human Siglec-2/CD22 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with human Siglec-2/CD22 and eGFP was stained with (A) Llama Anti-Human Siglec-2/CD22 Alexa Fluor® 594-conjugated Llamabody VHH Monoclonal Antibody (Catalog # LFAB10732T) or (B) normal llama IgG (IC011T). Staining was performed using our Staining Membrane-associated Proteins protocol.

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Preparation and Storage

Reconstitution
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Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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); 308501 (Rat); 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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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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