MPO, CD22 antibody

Catalog No. ABIN1112513

Product Details

Target: MPO, CD22

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Application: Flow Cytometry (FACS), Immunofluorescence (IF)

Characteristics: Monoclonal Mouse anti-human MPO, CD22. . The CD22 clone HIB22 monoclonal antibody is a heterodimer composed of two glycoprotein chains of 130 - 140 kDa molecular weight glycoprotein chains The antigen is present as both a cytoplasmic and membrane antigen on B lymphocytes. The CD22 antigen is expressed in the cytoplasm of the majority of B lymphocytes including early B progenitor cells. Antigen expression appears at an early stage in B lymphocyte differentiation at approximately the same stage as the CD19 antigen. Surface expression of the CD22 antigen by B lymphocytes is variable. Circulating B lymphocytes are CD22+ and follicular mantle and marginal zone B lymphocytes show strong binding of CBL147. Germinal centre B cells exhibit only weak expression of the CD22 antigen. The CD22 antigen is poorly expressed by non-T ALL cells and by a proportion of B- CLL cells. Hairy cell leukeamia cells have been demonstrated to strongly express the CD22 antigen.

Clone: 7-17-HIB22

Isotype: IgG1

Application Details

Application Notes: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence staining of human tissue for flow cytometric analysis using 20 µl/10^6 cells.

Sample Preparation:

Protocol for sample preparation and intracellular staining. 1. Pipette 50 µl of cell suspension to be analysed (up to 10^6 cells) into each tube. 2. For each sample, add an appropriate volume of conjugated antibody (20 µl) directed to the cell surface antigen of interest and the appropriate isotype control. Incubate for 15 minutes in the dark at room temperature. (This step is only necessary is you want to perform a direct immunofluorescence staining for a cell surface antigen) 3. Add 100 µL of Fix & Perm Reagent A, (Fixative), to each tube. Mix gently. 4. Incubate for 15 minutes at room temperature. 5. Wash once in 2 ml PBS Working Solution 1X. 6. Centrifuge for 5 minutes at 300 xg, then aspirate supernatant, leaving approximately 50 µl of fluid and vortex to ensure that the cell pellet are in suspension. 7. Add 100 µL of Fix & Perm Reagent B (Permeabilization), to each tube. Add the appropriate volume of conjugated intracellular antibody specific for the intracellular antigen and the appropriate isotype control. 8. Incubate for 15 minutes in the dark at room temperature. 9. Wash once in 2 ml PBS Working Solution 1X. 10. Centrifuge for 5 minutes at 300 xg, then aspirate supernatant, leaving approximately 50 µl of fluid and vortex to ensure that the cell pellet are in suspension. 11. Resuspend the cell pellet in 0,5 ml of 1% paraformaldehyde solution or an appropriate fluid for flow cytometric use, and store in the dark at 2-8 °C. Fixed cells should be analyzed within 24 hours.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: The conjugate is provided in liquid form in buffer containing 1% bovine serum albumin (BSA) and 0, 09% Sodium azide, pH 7.2. Clones: 7,17/ HIB22 Isotypes: IgG2a, IgG1 Flororchromes: Fluorescein isothiocyanate (Molecular Probes), R-Phycoerythrin (Europa Bioproducts, Ely, Cambridge).

Preservative: Sodium azide

Precaution of Use: 1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it must not be used in conjunction with patient records or treatment. 2. This product contains Sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, Sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.

Storage: 4 °C

Target Details

Target: MPO, CD22

Background: The MPO monoclonal antibody reacts with virtually all cells of the myeloid lineage (from myekloblast to neutrophils and monocytes). The monoclonal antibody does not react with human lymphocytes, thrombocytes and erythrocytes. 96% of all AML's are found positive, 100% of all ALL's are found negative.