Human Siglec-3/CD33 LlamabodyTM VHH His-tag Antibody
Human Siglec-3/CD33 LlamabodyTM VHH His-tag Antibody Summary
Accession # AAA51948
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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.
Detection of Siglec-3/CD33 in HEK293 Human Cell Line Transfected with Human Siglec-3/CD33 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either (A) human Siglec-3/CD33 or (B) irrelevant protein and eGFP was stained with Llama Anti-Human Siglec-3/CD33 Llamabody VHH Monoclonal Antibody (Catalog # LMAB10733) followed by Goat Anti-Llama Secondary Antibody (AF011), and then Allophycocyanin-conjugated Anti-Goat IgG Tertiary Antibody (F0108). Quadrant markers were set based on secondary plus tertiary antibody staining in the absence of primary antibody. View our protocol for Staining Membrane-associated Proteins.
Detection of Siglec‑3/CD33 in Transfected HEK293 Human Embryonic Kidney Cell Line (Positive) And Wild Type HEK293 Human Embryonic Kidney Cell Line (Negative) Cells. Siglec‑3/CD33 was detected in immersion fixed Transfected HEK293 Human Embryonic Kidney Cell Line (Positive) And Wild Type HEK293 Human Embryonic Kidney Cell Line (Negative) Cells using Llama Anti-Human Siglec‑3/CD33 LlamabodyTM VHH His-tag Monoclonal Antibody (Catalog # LMAB10733) at 5 µg/mL for 3 hours at room temperature. Cells were stained using Anti-camelid Vhh conjugated to Rhodamine-Red X (red) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Siglecs (sialic acid binding Ig-like lectins) are I-type (Ig-type) lectins belonging to the Ig superfamily. They are characterized by an N-terminal Ig-like V-type domain which mediates sialic acid binding, followed by varying numbers of Ig-like C2-type domains (1, 2). Eleven human Siglecs have been cloned and characterized. They are sialoadhesin/CD169/Siglec-1, CD22/Siglec-2, CD33/Siglec-3, Myelin-Associated Glycoprotein (MAG/Siglec-4a) and Siglecs 5 to 11 (1-3). To date, no Siglec has been shown to recognized any cell surface ligand other than sialic acids, suggesting that interactions with glycans containing this carbohydrate are important in mediating the biological functions of Siglecs. Siglecs 5 to 11 share a high degree of sequence similarity with CD33/Siglec-3 both in their extracellular and intracellular regions. They are collectively referred to as CD33-related Siglecs. One remarkable feature of the CD33-related Siglecs is their differential expression pattern within the hematopoietic system (1, 2). This fact, together with the presence of two conserved immunoreceptor tyrosine-based inhibition motifs (ITIMs) in their cytoplasma tails, suggests that CD33related Siglecs are involved in the regulation of cellular activation within the immune system. Human Siglec-3 is alternatively known as myeloid cell surface antigen CD33 and GP67. Human Siglec-3 cDNA encodes a 364 amino acid (aa) polypeptide with a hydrophobic signal peptide, an N-terminal Ig-like V-type domain, one Ig-like C2-type domains, a transmembrane region and a cytoplasmic tail (1, 4). Siglec-3 expression is restricted to cells of myelomonocytic lineage (2). It binds sialic acid preferring alpha 2,3-linkage over alpha 2,6-linkage (5). Studies indicated that Siglec-3 recruits SHP-1 and SHP-2 to its ITIMs (6, 7). When co‑cross‑linking with Fc gamma R1, Siglec3- inhibits tyrosine phosphorylation and calcium mobilization, suggesting Siglec-3 can mediate inhibitory signals (7).
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- Taylor, V.C. et al. (1999) J. Biol. Chem. 274:11505.
- Ulyanova, T. et al. (1999) Eur. J. Immunol. 29:3440.
Why is the Llamabody antibody called VHH His-tag? Does it detect His?
No, the antibody does not cross-react with His. It has a His-tag which is used for purification of the recombinant antibody.
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