Mouse anti-Human CD22 Antibody | anti-CD22 antibody

Mouse anti CD22, conjugated to PE

Cat. Num. AAA570369

• Gene Names: CD22; SIGLEC2; SIGLEC-2
• Reactivity: Human
• Applications: Flow Cytometry, Functional Assay, Immunofluorescence
• Synonyms: CD22; Mouse anti CD22; conjugated to PE; anti-CD22 antibody

• Host: Mouse
• Reactivity: Human
• Isotype: IgG1
• Clone Number: RFB4
• Specificity: The CD22 mAb (clone RFB4) recognizes surface CD22 expressed by mature peripheral B-cells and cytoplasmatic CD22 expressed by precursor B-cells.; ; The sensitivity of RFB4 mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI (mean fluorescence intensity).For this purpose, a mAb-concentration; range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 ul of leukocytes containing 10^7 cells/ml are stained with 20 ul mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
• Form/Format: Purified by Chromatography, Storage buffer: PBS pH 7.2, 1% BSA, 0.05% NaN3
• Sequence Length: 40

• Applicable Applications for anti-CD22 antibody: Flow Cytometry (FC/FACS), Indirect Immunofluorescence (IF)
• Application Notes: Staining Procedure for Surface CD22:; Direct Immunofluorescence (Staining Procedure) fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations; ; Proposed staining procedure for whole blood in short:; - For each sample add 50 ul of EDTA anti-coagulated blood to a 3-5 ml tube; - Add 20 ul of the appropriate monoclonal antibody conjugate; - Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark; - Add 100 ul to each tube and incubate for 10 minutes at room temperature - Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature; - Centrifuge tube for 5 minutes at 300 g; - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid; - Analyze immediately or store samples at 2-8 degree C in the dark and analyze within 24 hours; Proposed staining procedure for MNC in short:; - Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube; - Vortex at low speed for 1-2 seconds; - Incubate for 15-30 minutes at 2-8 degree C or at room temperature; - Centrifuge tubes for 5 minutes at 300 g; - Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g; - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8 degree C in the dark.; - Analyze fixed cells within 24 hours; ; Indirect Immunofluorescence (Staining Procedure); - Mix 20 ul purified antibody with 50 ul whole blood or MNC suspension; - Incubate for 15 minutes at 2-8 degree C; - Wash cells with phosphate buffered saline (PBS); - Add to cell pellet 20 ul of affinity purified, fluorochrome labeled F(ab')2 anti mouse Ig antibodies; - Incubate for 15 minutes at 2-8 degree C; - Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining; ; Staining Procedure for Cytoplasmatic CD22: Permeabilization and Staining Procedure; - In combination with our Permeabilization Kit FIX&PERM intracellular CD22 can be easily stained in cell suspensions.; - For each sample to be analyzed add 50 ul of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube; - Add 100 ul of Reagent A (Fixation Medium, stored and used at room temperature); - Incubate for 15 minutes at room temperature; - Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g; - Remove supernatant and add to cell pellet 100 ul Reagent B (Permeabilization Medium) and 20 ul of the CD22 monoclonal antibody conjugate; - Vortex at low speed for 1-2 seconds; - Incubate for 15 minutes at room temperature; - Wash cells with phosphate buffered saline as described above; - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1.0 % formaldehyde and store them at 2- 8 degree C in the dark.; - Analyze fixed cells within 24 hours.
• CE Mark: CE
• Conjugation: PE
• Preparation and Storage: Monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8 degree C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.

Related Product Information for anti-CD22 antibody

The epitope recognized by antibody RFB4 is a 135 kDa type I integral membrane protein expressed by human B-cells. Precursor B-cells are surface-CD22 negative, but cytoplasmic CD22 positive. Mature B-lymphocytes express CD22 also on their surface.

The RFB4 antibody permits the identification and enumeration of human B-cells using flow cytometry. Results must be put within the context of other tests as well
as the clinical history of the patient by a certified professional before final interpretation. Analysis performed with this antibody should be paralleled by positive and negative controls.

References

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NCBI and Uniprot Product Information

• NCBI GI #: 4126455
• NCBI GeneID: 933
• Molecular Weight: 75,354 Da
• NCBI Official Full Name: CD22, partial
• NCBI Official Synonym Full Names: CD22 molecule
• NCBI Official Symbol: CD22
• NCBI Official Synonym Symbols: SIGLEC2; SIGLEC-2
• NCBI Protein Information: B-cell receptor CD22; BL-CAM; CD22 antigen; T-cell surface antigen Leu-14; B-lymphocyte cell adhesion molecule; sialic acid binding Ig-like lectin 2; sialic acid-binding Ig-like lectin 2
• UniProt Protein Name: B-cell receptor CD22
• Protein Family: CD226 antigen
• UniProt Gene Name: CD22
• UniProt Synonym Gene Names: SIGLEC2; BL-CAM; Siglec-2
• UniProt Entry Name: CD22_HUMAN

Uniprot Description

CD22: Mediates B-cell B-cell interactions. May be involved in the localization of B-cells in lymphoid tissues. Binds sialylated glycoproteins; one of which is CD45. Preferentially binds to alpha-2,6-linked sialic acid. The sialic acid recognition site can be masked by cis interactions with sialic acids on the same cell surface. Upon ligand induced tyrosine phosphorylation in the immune response seems to be involved in regulation of B-cell antigen receptor signaling. Plays a role in positive regulation through interaction with Src family tyrosine kinases and may also act as an inhibitory receptor by recruiting cytoplasmic phosphatases via their SH2 domains that block signal transduction through dephosphorylation of signaling molecules. Belongs to the immunoglobulin superfamily. SIGLEC (sialic acid binding Ig-like lectin) family. 2 isoforms of the human protein are produced by alternative splicing.

Protein type: Membrane protein, integral; Cell surface

Chromosomal Location of Human Ortholog: 19q13.1

Cellular Component: integral to plasma membrane; external side of plasma membrane

Molecular Function: protein binding; carbohydrate binding

Biological Process: cell adhesion