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anti-Myeloperoxidase-C2 (FITC) and anti-CD3 (PE) antibody pair

ABIN1741603 产品详细信息, 供应商: Log in to see
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适用

宿主
小鼠
克隆类型 (克隆位点)
单克隆 ()
标记
FITC,PE
应用范围
Flow Cytometry (FACS)
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原理 This product is optimised for use with FIX&PERM®.
品牌 FIX&PERM®
克隆位点 8E6-UCHT1
亚型 IgG1
特异性 Antibody MPO-C2 (clone 8E2) reacts with human myeloperoxidase (MPO) expressed by normal and malignant myelomonocytic cells. The CD3 mAb (clone UCHT1) recognizes cytoplasmatic CD3 epsilon in precursor T-cells and cytoplasmic and surface CD3 epsilon in mature T-lymphocytes.In this COMBI-IC Reagent antibody 8E6 is conjugated to FITC, antibody UCHT1 is conjugated to Phycoeythrin (PE).
产品特性 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
组件 COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti CD3 (PE)
Sub Type Cocktail
分子量 14 kDa
应用备注 Permeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM- (ABIN1741575) intracellular MPO-C2 and 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 MPO-C2/CD3 COMBI-IC 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
说明

Myeloperoxidase (MPO) is a glycoprotein present in the azurophil (primary) granules of myeloid cells, which appears in the myeloblast stage of myeloid cell differentiation. MPO is he most common functional protein of myeloid cells and is involved in the inflammatory response. It helps to kill microbes by breaking down peroxide in the presence of halide ions, contributing to the bactericidal function of granulocytes. The primary translation product of MPO undergoes glycosylation with production of the 89 kDa heme-free apopro-MPO form followed by incorporation of heme and conversion into the enzymatically active pro-MPO form. Subsequently, pro-MPO becomes targeted to azurophil granules where final processing occurs to produce mature dimeric MPO consisting of the 59-64 kDa MPO a-chain and the 14 kDa MPO b-chain. Precursor T-cells are surface CD3 negative but positive for cytoplasmatic CD3. All mature T cells are both cytoplasmic and surface CD3 positive. The combined staining for MPO and CD3 allows the distinction of cells derived form the myeloid lineage that are generally MPO positive, from immature and mature T lymphocytes during normal and malignant hematopoiesis. The MPO-C2/CD3 COMBI-IC reagent permits the identification and enumeration of normal and malignant 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.

实验流程

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
缓冲液 PBS pH 7.2, 1 mg/mL BSA, 0.0.5 % sodium azide
储存液 Sodium azide
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.
注意事项 Do not freeze and protect from prolonged exposure to light.
储存条件 4 °C
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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Nauseef: "Myeloperoxidase deficiency." in: Hematologic pathology, Vol. 4, Issue 4, pp. 165-78, 1991 (PubMed).

Srivastava, Rado, Bauerle, Broxmeyer: "Regulation of human bone marrow lactoferrin and myeloperoxidase gene expression by tumor necrosis factor-alpha." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 146, Issue 3, pp. 1014-9, 1991 (PubMed).

Catovsky, Matutes, Buccheri, Shetty, Hanslip, Yoshida, Morilla: "A classification of acute leukaemia for the 1990s." in: Annals of hematology, Vol. 62, Issue 1, pp. 16-21, 1991 (PubMed).

van der Schoot, Daams, Pinkster, Vet, von dem Borne: "Monoclonal antibodies against myeloperoxidase are valuable immunological reagents for the diagnosis of acute myeloid leukaemia." in: British journal of haematology, Vol. 74, Issue 2, pp. 173-8, 1990 (PubMed).

Janossy, Coustan-Smith, Campana: "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).

Rani, De Oliveira, Catovsky: "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).

Vainchenker, Villeval, Tabilio, Matamis, Karianakis, Guichard, Henri, Vernant, Rochant, Breton-Gorius: "Immunophenotype of leukemic blasts with small peroxidase-positive granules detected by electron microscopy." in: Leukemia, Vol. 2, Issue 5, pp. 274-81, 1988 (PubMed).

Nauseef, Olsson, Arnljots: "Biosynthesis and processing of myeloperoxidase--a marker for myeloid cell differentiation." in: European journal of haematology, Vol. 40, Issue 2, pp. 97-110, 1988 (PubMed).

van der Schoot, von dem Borne, Tetteroo: "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).

Murao, Stevens, Ito, Huberman: "Myeloperoxidase: a myeloid cell nuclear antigen with DNA-binding properties." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, Issue 4, pp. 1232-6, 1988 (PubMed).

Campana, Thompson, Amlot, Brown, Janossy: "The cytoplasmic expression of CD3 antigens in normal and malignant cells of the T lymphoid lineage." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 138, Issue 2, pp. 648-55, 1987 (PubMed).

Koeffler, Ranyard, Pertcheck: "Myeloperoxidase: its structure and expression during myeloid differentiation." in: Blood, Vol. 65, Issue 2, pp. 484-91, 1985 (PubMed).