Mouse anti CD3

• Catalog number
  GM-4011
• Price
  Please ask
• Size
  0,2mg

• Category
  Primary Antibodies
• Long description
  UCHT1 is directed against human CD3 – the multichain complex associated with the T-cell receptor. Precursor T-cells are surface CD3 negative but positive for cytoplasmic CD3. All mature T-cells are both cytoplasmic and surface CD3 positive. The UCHT1 antibody permits the identification and enumeration of normal and leukemic human blood and bone marrow 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
  UCHT1
• Antigen antibody binding interaction
  Mouse anti CD3 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 CD3 mAb (clone UCHT1) recognizes cytoplasmic CD3 epsilon in precursor T-cells and cytoplasmic and surface CD3 epsilon in mature T-lymphocytes. The sensitivity of UCHT1 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).The sensitivity of UCHT1 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).In practice, 50 µl of leukocytes containing 107 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 CD3: 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 CD3: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM® (Cat. No. GAS-002) intracellular CD3 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 CD3 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. Catovsky, D., Matutes, E., Buccheri, V., Shetty, V., Hanslip, J., Yoshida, N. & Morilla, R. (1991) Ann Hematol 62, 16-21. Janossy, G., Coustan-Smith, E. & Campana, D. (1989) Leukemia 3, 170-81. Clevers, H., Alarcon, B., Wileman, T. & Terhorst, C. (1988) Annu Rev Immunol 6, 629-62. Wering, E. R. & Terhorst, C. (1988) Blood 71, 603-12. Rani, S., De Oliveira, M. S. & Catovsky, D. (1988) Hematol Pathol 2, 73-8. van der Schoot, C. E., von dem Borne, A. E. & Tetteroo, P. A. (1987) Acta Haematol 78 Suppl 1, 32-40. van Dongen, J. J., Krissansen, G. W., Wolvers-Tettero, I. L., Comans-Bitter, W. M., Adriaansen, H. J., Hooijkaas, H., van Campana, D., Thompson, J. S., Amlot, P., Brown, S. & Janossy, G. (1987) J Immunol 138, 648-55. Burns, G. F., Boyd, A. W. & Beverley, P. C. (1982) J Immunol 129, 1451-7 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
  CD3  
• Gene symbol
  CD3G, CD3E
• Short name
  Mouse anti CD3
• Technique
  Mouse, anti, antibody to, mouses
• Host
  mouse
• Isotype
  IgG1
• Label
  unlabeled
• Species
  Mouse, Mouses
• Alternative name
  Mouse antibody to CD3
• Alternative technique
  murine, antibodies

Gene info

• Identity
  HGNC:1675
• Gene
  CD3G
• Long gene name
  CD3g molecule
• Synonyms gene name
  • CD3g antigen, gamma polypeptide (TiT3 complex)
  • CD3g molecule, gamma (CD3-TCR complex)
• GenBank acession
  • X60491
• Locus
  11q23.3
• Discovery year
  1988-05-11
• Entrez gene record
  917
• RefSeq identity
  • NM_000073
• Classification
  • CD molecules
• VEGA ID
  OTTHUMG00000166971
• Locus Specific Databases
  • CD3Gbase: Mutation registry for autosomal recessiveCD3gamma immunodeficiency
  • LRG_39

Gene info

• Identity
  HGNC:1674
• Gene
  CD3E
• Long gene name
  CD3e molecule
• Synonyms gene name
  • CD3e antigen, epsilon polypeptide (TiT3 complex)
  • CD3e molecule, epsilon (CD3-TCR complex)
• GenBank acession
  • X03884
• Locus
  11q23.3
• Discovery year
  1986-01-01
• Entrez gene record
  916
• RefSeq identity
  • NM_000733
• Classification
  • CD molecules
• VEGA ID
  OTTHUMG00000166968
• Locus Specific Databases
  • CD3Ebase: Mutation registry for autosomal recessive CD3epsilon immunodeficiency
  • LRG_38

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

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