Anti-Mouse CD22 (Clone MB22-11) – Purified in vivo GOLD™ Functional Grade

Anti-Mouse CD22 (Clone MB22-11) – Purified in vivo GOLD™ Functional Grade

Product No.: C960

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Clone
MB22-11
Target
CD22
Formats AvailableView All
Product Type
Hybridoma Monoclonal Antibody
Alternate Names
Lyb-8, Siglec-2, BL-CAM
Isotype
Mouse IgG2c κ
Applications
ELISA
,
FA

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Mouse
Immunogen
Mouse CD22 cDNA-transfected baby hamster kidney cells
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
≤ 1.0 EU/mg as determined by the LAL method
Purity
≥95% by SDS Page
≥95% monomer by analytical SEC
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
State of Matter
Liquid
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using only in vitro protein free cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at -70°C. Avoid Repeated Freeze Thaw Cycles.
Regulatory Status
Research Use Only
Country of Origin
USA
Shipping
2-8°C Wet Ice
Additional Applications Reported In Literature ?
ELISA,
FA
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Specificity
MB22-11 activity is directed against mouse CD22 (Siglec-2).
Background
Siglecs (sialic acid-binding immunoglobulin superfamily lectins) are a family of single pass, transmembrane cell surface proteins characterized by shared structural motifs and an ability to recognize sialic acids1, 2. CD22 (Siglec-2), a 140 kDa member of the Siglec family expressed by B cells3, 4, contains six C2-set domains, one V-set domain, and in its intracellular cytoplasmic tail has three immunoreceptor tyrosine-based inhibition motifs (ITIM) and one ITIM-like domain5. While murine Siglecs are not necessarily homologous to human Siglecs, CD22 is evolutionarily conserved and does have a direct human ortholog5.

CD22 acts as an inhibitory B cell co-receptor that negatively regulates B cell activation, B reg cell expansion, and B cell receptor (BCR) signaling4. Upon ligation of BCR, ITIMs are phosphorylated, leading to recruitment and activation of SH2-containing phosphatases that then dephosphorylate signaling molecules activated by BCR ligation4. Additionally, CD22 regulates B cell response to inflammation and is a master regulator of microglial phagocytosis in the aging brain5.

Evidence in mouse models suggests CD22 contributes to the pathogenesis of autoimmune diseases3. Loss of CD22 leads to hyperactivation of B cells5. CD22 mouse knockouts are defective in B cell development but do not develop lupus-like disease4.

To generate MB22-11, CD22 knockout mice were immunized with mouse CD22 cDNA-transfected baby hamster kidney cells6. Spleen cells were fused with NS-1 myeloma cells, and hybridomas producing antibody specifically reactive with CD22-transfected mouse L cells were selected and purified. MB22-11 was isotyped as IgG2c due to its C57BL/6 origin; however, both IgG2a and IgG2c specific reagents have significant reactivity against MB22-11.

In vitro, MB22-11 inhibits CD22-mediated adhesion by 90% and completely blocks CD22-Fc binding to T and B cells6. In vivo, MB22-11 significantly reduces peripheral blood, lymph node, and marginal zone B cell numbers6, 7. Additionally, in mice injected with MB22-11, blood, spleen, and lymph node B cell turnover is higher relative to injection with non-blocking monoclonal antibodies, and B cell surface expression of CD22 is reduced to nearly undetectable levels6.
Antigen Distribution
CD22 is expressed by most mature B cell lineages.

Antigen Details

Ligand/Receptor
SHP-1, Syk, Lck, and Lyn
NCBI Gene Bank ID
UniProt.org
Research Area
Cell Adhesion
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Immunology

References & Citations

1. Bochner BS. Clin Exp Allergy. 39(3):317-324. 2009.
2. Kiwamoto T, Kawasaki N, Paulson JC, et al. Pharmacol Ther. 135(3):327-336. 2012.
3. Dörner T, Shock A, Smith KG. Int Rev Immunol. 31(5):363-378. 2012.
4. Tsubata T. Immunol Med. 42(3):108-116. 2019.
5. Siddiqui SS, Matar R, Merheb M, et al. Cells. 8(10):1125. 2019.
6. Haas KM, Sen S, Sanford IG, et al. J Immunol. 177(5):3063-3073. 2006.
7. Haas KM, Watanabe R, Matsushita T, et al. J Immunol. 184(9):4789-4800. 2010.
Indirect Elisa Protocol
FA

Certificate of Analysis

Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.