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Mouse Siglec-2/CD22 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF2296

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AF2296-SP
AF2296

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Siglec‑2/CD22
Ser22-Arg702
Accession # P35329

Specificity

Detects mouse Siglec‑2/CD22 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross‑reactivity with recombinant human (rh) Siglec-2 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse Siglec-2/CD22 Antibody

Cell Adhesion Mediated by Siglec‑2/CD22 and Neutral-ization by Mouse Siglec‑2/ CD22 Antibody.

Cell Adhesion Mediated by Siglec‑2/CD22 and Neutral-ization by Mouse Siglec‑2/ CD22 Antibody.

Recombinant Mouse Siglec-2/ CD22 Fc Chimera (Catalog # 2296-SL), immobilized onto a microplate, supports the adhesion of human red blood cells in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Mouse Siglec-2/ CD22 Fc Chimera (0.3 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse Siglec-2/CD22 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2296). The ND50 is typically 0.1-0.5 µg/mL.
Siglec-2/CD22 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

Siglec‑2/CD22 in Mouse Splenocytes.

Siglec-2/CD22 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse Siglec-2/CD22 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2296) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Siglec-2/CD22 by Flow Cytometry

Detection of Siglec-2/CD22 by Flow Cytometry

High-protein diet reduces villus-resident CD22+ eosinophils. (A) Percent of kilocalories (% kCal) from different macronutrients for diets shown in B. RP = reduced (8%) protein, C = control (18% protein), HP = high (70%) protein, and HF = high fat. (B) Eosinophil frequency in the proximal SI lamina propria (LP) of mice fed diets containing variable macronutrient composition for 15 d, representative of at least 3 experiments with 3 to 4 mice per group. (C and D) Eosinophil frequency (C) and number (D) in the proximal SI LP of mice fed diets containing the indicated percentage of protein (15 d), representative of 2 experiments with 4 to 6 mice per group. (E) Eosinophil frequency in proximal SI LP of mice fed either C, HP, or a diet containing elevated amino acids (“HAA”) in amounts equivalent to those in HP (15 d), representative of 2 experiments with 4 mice per group. (F and G) Representative microscopy images (F) and quantification (G) of eosinophils near crypts and in villi, per region, of the proximal SI of mice fed C or HP diets [red = MBP, blue = DAPI, (Scale bar, 100 μm.) “Duod.” = duodenum, “Jejun.” = jejunum]. (H) Eosinophil frequency in LP of the indicated regions of the gastrointestinal (GI) tract in mice fed C or HP diets for 15 d, representative of 2 experiments with 4 mice/group. (I) Percent of eosinophils that were CD22+ in LP of the indicated regions of the GI tract in mice fed C or HP diet for 15 d, representative of 2 experiments with 4 mice per group. (J) Representative flow cytometry plots showing alpha4 beta7 and CD22 staining in proximal SI LP eosinophils in mice fed C or HP diet for 15 d. Data are presented as mean ± SEM. All data were analyzed by Student’s t test. ∗∗∗∗P < 0.0001, ∗∗∗P < 0.001, ∗∗P < 0.01, ∗P < 0.05, and ns = not significant (P > 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38271336), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse Siglec-2/CD22 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse Siglec-2/CD22 Fc Chimera (Catalog # 2296-SL)

Neutralization

Measured by its ability to neutralize Siglec-2/CD22-mediated adhesion of human red blood cells. Kelm, S. et al. (1994) Current Biology 4:965. The Neutralization Dose (ND50) is typically 0.1-0.5 µg/mL in the presence of 0.3 µg/mL Recombinant Mouse Siglec-2/CD22 Fc Chimera.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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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-2/CD22

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. Among these are sialoadhesin/CD169/Siglec-1, CD22/Siglec-2 and CD33/Siglec-3. To date, no Siglec has been shown to recognize any cell surface ligand other than sialic acid, suggesting that interactions with glycans containing this carbohydrate are important in mediating the biological functions of Siglecs. The cDNA of mouse Siglec-2 (also known as B-cell antigen CD22), encodes an 862 amino acid (aa) protein that contains a 21 aa signal peptide, a 681 aa extracellular region, a 19 aa transmembrane region and a 141 aa cytoplasmic tail (3, 4). The extracellular region contains one N-terminal V-type Ig-like domain followed by six Ig-like C2-type domains. The cytoplasmic domain has 3 immunoreceptor tyrosine-based inhibition motifs (ITIMs). Two splice forms exist, both showing deletions in the V-type Ig domain of 30 aa and 60 aa each. There are also two alleles in mouse that account for a difference of 10 aa in the extracellular region. The extracellular region of mouse Siglec-2 is 60% aa identity to human extracellular Siglec-2. Expression of mouse Siglec-2/CD22 generates a 140 kDa integral membrane glycoprotein that is limited to the B cell compartment of lymphoid tissues. Its expression is upregulated by LPS activtion (5, 6). Siglec-2/CD22 is an adhesion molecule that preferentially binds alpha2,6- 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 (7).

References

  1. Nitschke, L. et al. (2001) Scand. J. Immunol. 53:227.
  2. Crocker, P.R. and A. Varki (2001) Immunology 103:137.
  3. Law, C-L. et al. (1993) J. Immunol. 151:175.
  4. Wienands, Y.J. et al. (1999) J. Biol. Chem. 274:18769.
  5. Wilson, G.L et al. (1991) J. Exp. Med. 173:137.
  6. Torres, P.M. (1992) J. Immunol. 194:2641.
  7. Collins, B.E. et al. (2004) Proc. Natl. Acad. Sci. 101:6104.

Long Name

Sialic Acid Binding Ig-like Lectin 2

Alternate Names

BL-CAM, CD22, Siglec2

Entrez Gene IDs

933 (Human); 12483 (Mouse); 308501 (Rat); 102130968 (Cynomolgus Monkey)

Gene Symbol

CD22

UniProt

Additional Siglec-2/CD22 Products

Product Documents for Mouse Siglec-2/CD22 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse Siglec-2/CD22 Antibody

For research use only

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