Mouse anti CD22

• Catalog number
  GM-4051
• Price
  Please ask
• Size
  0,2mg

• Category
  Primary Antibodies
• Long description
  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 diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us.
• Antibody come from
  n/a
• Other description
  Purified by Chromatography, Storage buffer: PBS pH 7.2, 1% BSA, 0.05% NaN3
• Clone
  RFB4
• Antigen antibody binding interaction
  Mouse anti CD22 Antibody
• Antibody is raised in
  Mouse
• Antibody s reacts with
  Human
• Antibody s reacts with these species
  This antibody doesn't cross react with other species
• Antibody s 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 µl of leukocytes containing 10^7 cells/ml are stained with 20 µl 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.
• Research interest
  CD Marker
• Application
  Flow cytometry, Indirect Immunofluorescence
• Antibody s suited for
  Staining Procedure for Surface CD22: Direct Immunofluorescence (Staining Procedure) Nordic-MUbio 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 µl of EDTA anti-coagulated blood to a 3-5 ml tube - Add 20 µl of the appropriate Nordic-MUbio monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4°C or at room temperature in the dark - Add 100 µl Nordic-MUbio-LYSE (Cat.No. GAS-003) 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° C in the dark and analyze within 24 hours For “No-Wash” protocol please refer to www.nordicmubio.com Proposed staining procedure for MNC in short: - Carefully add 20 µl antibody conjugate and 50-100 µl MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8°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°C in the dark. - Analyze fixed cells within 24 hours Indirect Immunofluorescence (Staining Procedure) - Mix 20 µl Nordic-MUbio purified antibody with 50 µl whole blood or MNC suspension - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS) - Add to cell pellet 20 µl of affinity purified, fluorochrome labeled F(ab’)2 anti mouse Ig antibodies - Incubate for 15 minutes at 2-8°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® (Cat. No. GAS-002) intracellular CD22 can be easily stained in cell suspensions. - For each sample to be analyzed add 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube - Add 100 µl 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 µl Reagent B (Permeabilization Medium) and 20 µl 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°C in the dark. - Analyze fixed cells within 24 hours.
• Storage
  Nordic-MUbio 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°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.
• Relevant references
  Paietta, E. (2003) Best Pract Res Clin Haematol 16, 671-83. Braylan, R. C., Orfao, A., Borowitz, M. J. & Davis, B. H. (2001) Cytometry 46, 23-7. Lanza, F., Latorraca, A., Moretti, S., Castagnari, B., Ferrari, L. & Castoldi, G. (1997) Cytometry 30, 134-44. Groeneveld, K., te Marvelde, J. G., van den Beemd, M. W., Hooijkaas, H. & van Dongen, J. J. (1996) Leukemia 10, 1383-9. Wilson, G. L., Fox, C. H., Fauci, A. S. & Kehrl, J. H. (1991) J Exp Med 173, 137-46. Catovsky, D., Matutes, E., Buccheri, V., Shetty, V., Hanslip, J., Yoshida, N. & Morilla, R. (1991) Ann Hematol 62, 16-21. Stamenkovic, I. & Seed, B. (1990) Nature 345, 74-7. Janossy, G., Coustan-Smith, E. & Campana, D. (1989) Leukemia 3, 170-81. Li, J. L., Shen, G. L., Ghetie, M. A., May, R. D., Till, M., Ghetie, V., Uhr, J. W., Janossy, G., Thorpe, P. E., Amlot, P. & et al. (1989) Cell Immunol 118, 85-99. Schwartz-Albiez, R., Dorken, B. & Moldenhauer, G. (1989) In Leukocyte Typing IV (Oxford University Press, Oxford) p65-7. Pezzutto, A., Rabinovitch, P. S., Dorken, B., Moldenhauer, G. & Clark, E. A. (1988) J Immunol 140, 1791-5. Rani, S., De Oliveira, M. S. & Catovsky, D. (1988) Hematol Pathol 2, 73-8. Boue, D. R. & Lebien, T. W. (1988) J Immunol 140, 192-9. Mason, D. Y., Stein, H., Gerdes, J., Pulford, K. A., Ralfkiaer, E., Falini, B., Erber, W. N., Micklem, K. & Gatter, K. C. (1987) Blood 69, 836-40. Pezzutto, A., Dorken, B., Moldenhauer, G. & Clark, E. A. (1987) J Immunol 138, 98-103. van der Schoot, C. E., von dem Borne, A. E. & Tetteroo, P. A. (1987) Acta Haematol 78 Suppl 1, 32-40. Dorken, B., Moldenhauer, G., Pezzutto, A., Schwartz, R., Feller, A., Kiesel, S. & Nadler, L. M. (1986) J Immunol 136, 4470-9. Beverley, P. C., Linch, D. & Callard, R. E. (1981) Haematol Blood Transfus 26, 309-13.
• Protein number
  see ncbi
• Warnings
  For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.
• Antibody s link
• Description
  This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided.
• Test
  Mouse or mice from the Mus musculus species are used for production of mouse monoclonal antibodies or mabs and as research model for humans in your lab. Mouse are mature after 40 days for females and 55 days for males. The female mice are pregnant only 20 days and can give birth to 10 litters of 6-8 mice a year. Transgenic, knock-out, congenic and inbread strains are known for C57BL/6, A/J, BALB/c, SCID while the CD-1 is outbred as strain.
• Latin name
  Mus musculus

• Gene target
  CD22  
• Gene symbol
  CD22
• Short name
  Mouse anti CD22
• Technique
  Mouse, anti, antibody to, mouses
• Host
  mouse
• Isotype
  IgG1
• Label
  unlabeled
• Species
  Mouse, Mouses
• Alternative name
  Mouse antibody to CD22 molecule
• Alternative technique
  murine, antibodies
• Alternative to gene target
  CD22 molecule, SIGLEC-2 and SIGLEC2, CD22 and IDBG-44001 and ENSG00000012124 and 933, protein binding, Cell surfaces, Cd22 and IDBG-171574 and ENSMUSG00000030577 and 12483, CD22 and IDBG-635610 and ENSBTAG00000011887 and 514582

Gene info

• Identity
  HGNC:1643
• Gene
  CD22
• Long gene name
  CD22 molecule
• Synonyms gene name
  • CD22 antigen
• Synonyms
  • SIGLEC-2
  • SIGLEC2
• Synonyms name
  • sialic acid binding Ig-like lectin 2
• GenBank acession
  • X52785
• Locus
  19q13.12
• Discovery year
  1991-06-06
• Entrez gene record
  933
• Pubmed identfication
  • 8496602
  • 1691828
• RefSeq identity
  • NM_001771
• Classification
  • CD molecules
  • MicroRNA protein coding host genes
  • C2-set domain containing
  • Sialic acid binding Ig like lectins
• VEGA ID
  OTTHUMG00000183298

MeSH Data

• Name
  Blotting, Western
• Concept
  Scope note: Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
• Tree numbers
  • E05.196.401.143
  • E05.301.300.096
  • E05.478.566.320.200
  • E05.601.262
  • E05.601.470.320.200
• Qualifiers
  ethics, trends, veterinary, history, classification, economics, instrumentation, methods, standards, statistics & numerical data

Product images

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