Why is the Llamabody antibody called VHH His-tag? Does it detect His?

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 Llamabody^TM 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.

Applications for Human Siglec-3/CD33 Llamabody^TM VHH His-tag Antibody

Application

Recommended Usage

Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 Human Cell Line Transfected with Human Siglec-3/CD33 and eGFP

Immunocytochemistry

5-25 µg/mL
Sample: Immersion fixed Transfected HEK293 Human Embryonic Kidney Cell Line (Positive) And Wild Type HEK293 Human Embryonic Kidney Cell Line (Negative) Cells

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

His-tag purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Siglec-3/CD33

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 alpha2,3-linkage over alpha2,6-linkage (5). Studies indicated that Siglec-3 recruits SHP-1 and SHP-2 to its ITIMs (6, 7). When co-cross-linking with Fc gammaR1, Siglec3- inhibits tyrosine phosphorylation and calcium mobilization, suggesting Siglec-3 can mediate inhibitory signals (7).

References

Crocker, P.R. and A. Varki (2001) Trends Immunol. 22:337.
Crocker, P.R. and A. Varki (2001) Immunology 103:137.
Angata, T. et al. (2002) J. Biol. Chem. 277:24466.
Simmons, D. and B. Seed (1988) J. Immunol. 141:2797.
Freeman, S.D. et al. (1995) Blood 85:2002.
Taylor, V.C. et al. (1999) J. Biol. Chem. 274:11505.
Ulyanova, T. et al. (1999) Eur. J. Immunol. 29:3440.

Long Name

Sialic Acid Binding Ig-like Lectin 3

Alternate Names

CD33, gp67, Siglec3

Entrez Gene IDs

945 (Human); 12489 (Mouse); 102117580 (Cynomolgus Monkey)

Gene Symbol

CD33

UniProt

AAA51948

Additional Siglec-3/CD33 Products

Product Documents

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COA

SDS

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Product Specific Notices for Human Siglec-3/CD33 Llamabody^TM VHH His-tag Antibody

For research use only

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Flow Cytometry

Human Siglec-3/CD33 LlamabodyTM VHH His-tag Antibody

R&D Systems, part of Bio-Techne | Catalog # LMAB10733

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

View all Siglec-3/CD33 Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Siglec-3/CD33 LlamabodyTM VHH His-tag Antibody

Detection of Siglec-3/CD33 in HEK293 Human Cell Line Transfected with Human Siglec-3/CD33 and eGFP by Flow Cytometry.