Mouse anti-Human CD3 Antibody | anti-CD3 antibody
Mouse anti CD3, conjugated to PE
Reactivity
Human
Applications
Flow Cytometry, Functional Assay, Immunofluorescence
Synonyms
CD3; Mouse anti CD3; conjugated to PE; anti-CD3 antibody
Host
Mouse
Reactivity
Human
Isotype
IgG1
Clone Number
UCHT1
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 ul of leukocytes containing 107 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
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 ul of leukocytes containing 107 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
167
Applicable Applications for anti-CD3 antibody
Flow Cytometry (FC/FACS), Indirect Immunofluorescence (IF)
Application Notes
Staining Procedure for Surface CD3:
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 CD3: Permeabilization and Staining Procedure
- In combination with our Permeabilization Kit FIX&PERM intracellular CD3 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 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 degree C in the dark.
- Analyze fixed cells within 24 hours.
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 CD3: Permeabilization and Staining Procedure
- In combination with our Permeabilization Kit FIX&PERM intracellular CD3 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 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 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-CD3 antibody
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 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.
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 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
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.
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.
NCBI and Uniprot Product Information
NCBI GI #
Molecular Weight
18,897 Da
NCBI Official Full Name
cd3
UniProt Protein Name
CD3gamma/delta
Protein Family
UniProt Gene Name
cd3g
UniProt Entry Name
P79951_XENLA
Similar Products
Product Notes
The CD3 cd3g (Catalog #AAA570365) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Mouse anti CD3, conjugated to PE reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's CD3 can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS), Indirect Immunofluorescence (IF). Staining Procedure for Surface CD3: 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 CD3: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM intracellular CD3 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 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 degree C in the dark. - Analyze fixed cells within 24 hours. . Researchers should empirically determine the suitability of the CD3 cd3g for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD3, Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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