CD3 抗体 (FITC)
Quick Overview for CD3 抗体 (FITC) (ABIN1741588)
抗原
See all CD3 抗体适用
宿主
克隆类型
标记
应用范围
克隆位点
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原理
- This product is optimised for use with FIX&PERM®.
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品牌
- FIX&PERM®
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特异性
- 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
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产品特性
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Mildly fixes cells, preserving their flow cytometric scatter characteristics
Allows simultaneous characterisation of both intracellular and cell surface markers
Rapid technique - whole procedure can be carried out in less than one hour, ready
for immediate analysis or storage for 24 hours
Stringent QC procedures - the quality of each lot is determined using well-defined
blood samples and subsequent comparison of scatter characteristics of obtained
leukocyte populations, ensuring consistent and reliable results lot after lot
A range of intracellular antibodies with optimised protocols for use with FIX&PERM®
FIX&PERM® is a simple procedure making use of two reagents. Reagent A gently fixes cells, while Reagent B permeabilizes them. The specific formulations reduce background and allow simultaneous addition of permeabilization medium and fluorochrome labelled antibodies, allowing staining of intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins, etc. -
纯化方法
- Purified by Affinity Chromatography
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亚型
- IgG1
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应用备注
- 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) preparationsProposed 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 monoclonal antibody conjugate - Incubate the tube for 15 minutes at 4°C or at room temperature in the dark - Add 100 µL -LYSE (ABIN1741580) 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 hoursFor \No-Wash\ 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 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 stainingStaining Procedure for Cytoplasmatic CD3: Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM® (ABIN1741575) 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 5 mL tube- Add 100 µL of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5 mL 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.
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说明
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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.
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实验流程
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Procedure: For each sample to be analysed use 50 µL of whole blood, bone marrow or mononuclear cell suspension in a 5 mL tube
Add 100 µL of Reagent A (Fixation Medium, stored and used at room temperature)
Incubate for 15 minutes at room temperature
Add 5 mL 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 appropriate 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 re-suspend cells in sheath fluid for immediate analysis or re-suspend cells in 0.5 mL 1.0 % formaldehyde and store at 2-8°C in the dark
Analyse fixed cells within 24 hours -
限制
- 仅限研究用
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状态
- Liquid
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缓冲液
- PBS pH 7.2, 1 % BSA, 0.05 % sodium azide
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储存液
- Sodium azide
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注意事项
- 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.
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注意事项
- Do not freeze and protect from prolonged exposure to light.
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储存条件
- 4 °C
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储存方法
- These reagents should be stored at 2-8°C 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.
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: "Two phase 3 trials of bapineuzumab in mild-to-moderate Alzheimer's disease." in: The New England journal of medicine, Vol. 370, Issue 4, pp. 322-33, (2014) (PubMed).
: "Optimal number of reagents required to evaluate hematolymphoid neoplasias: results of an international consensus meeting." in: Cytometry, Vol. 46, Issue 1, pp. 23-7, (2001) (PubMed).
: "Comparative analysis of different permeabilization methods for the flow cytometry measurement of cytoplasmic myeloperoxidase and lysozyme in normal and leukemic cells." in: Cytometry, Vol. 30, Issue 3, pp. 134-44, (1997) (PubMed).
: "Flow cytometric detection of intracellular antigens for immunophenotyping of normal and malignant leukocytes." in: Leukemia, Vol. 10, Issue 8, pp. 1383-9, (1996) (PubMed).
: "A classification of acute leukaemia for the 1990s." in: Annals of hematology, Vol. 62, Issue 1, pp. 16-21, (1991) (PubMed).
: "The reliability of cytoplasmic CD3 and CD22 antigen expression in the immunodiagnosis of acute leukemia: a study of 500 cases." in: Leukemia, Vol. 3, Issue 3, pp. 170-81, (1989) (PubMed).
: "Different expression of CD3 and CD22 in leukemic cells according to whether tested in suspension or fixed on slides." in: Hematologic pathology, Vol. 2, Issue 2, pp. 73-8, (1989) (PubMed).
: "The T cell receptor/CD3 complex: a dynamic protein ensemble." in: Annual review of immunology, Vol. 6, pp. 629-62, (1988) (PubMed).
: "Characterization of myeloid leukemia by monoclonal antibodies, with an emphasis on antibodies against myeloperoxidase." in: Acta haematologica, Vol. 78 Suppl 1, pp. 32-40, (1988) (PubMed).
: "Cytoplasmic expression of the CD3 antigen as a diagnostic marker for immature T-cell malignancies." in: Blood, Vol. 71, Issue 3, pp. 603-12, (1988) (PubMed).
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: "Two phase 3 trials of bapineuzumab in mild-to-moderate Alzheimer's disease." in: The New England journal of medicine, Vol. 370, Issue 4, pp. 322-33, (2014) (PubMed).
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- CD3
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别名
- CD3
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途径
- TCR Signaling, Ubiquitin Proteasome Pathway
抗原
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