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Flow cytometryAntibody data
- Antibody Data
- Antigen structure
- References [9]
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- Validations
- Flow cytometry [1]
- Other assay [4]
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- Product number
- 47-1239-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD123 Monoclonal Antibody (6H6), APC-eFluor™ 780, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The 6H6 monoclonal antibody reacts with human CD123, the alpha chain of the IL-3 receptor. This 60-70 kDa transmembrane protein binds to IL-3 with low affinity by itself, and when associated with CD131 (common beta chain) binds IL-3 with high affinity. CD123 is expressed by myeloid precursors, macrophages, dendritic cells, mast cells, basophils, and megakaryocytes. Applications Reported: This 6H6 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 6H6 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. APC-eFluor® 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 633-647 nm; Emission: 780 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 6H6
- Vial size
- 25 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Unsupervised Analysis of Flow Cytometry Data in a Clinical Setting Captures Cell Diversity and Allows Population Discovery.
Neutrophils Inhibit Synthesis of Mineralized Extracellular Matrix by Human Bone Marrow-Derived Stromal Cells In Vitro.
Malaria-induced NLRP12/NLRP3-dependent caspase-1 activation mediates inflammation and hypersensitivity to bacterial superinfection.
Frailty in old age is associated with decreased interleukin-12/23 production in response to toll-like receptor ligation.
CD56-positive haematological neoplasms of the skin: a multicentre study of the Cutaneous Lymphoma Project Group of the European Organisation for Research and Treatment of Cancer.
Epithelial cells trigger frontline immunoglobulin class switching through a pathway regulated by the inhibitor SLPI.
Local activation of the innate immune system in Buruli ulcer lesions.
Expression of the plasmacytoid dendritic cell marker BDCA-2 supports a spectrum of maturation among CD4+ CD56+ hematodermic neoplasms.
TCL1 expression in plasmacytoid dendritic cells (DC2s) and the related CD4+ CD56+ blastic tumors of skin.
Baumgaertner P, Sankar M, Herrera F, Benedetti F, Barras D, Thierry AC, Dangaj D, Kandalaft LE, Coukos G, Xenarios I, Guex N, Harari A
Frontiers in immunology 2021;12:633910
Frontiers in immunology 2021;12:633910
Neutrophils Inhibit Synthesis of Mineralized Extracellular Matrix by Human Bone Marrow-Derived Stromal Cells In Vitro.
Bastian OW, Croes M, Alblas J, Koenderman L, Leenen LPH, Blokhuis TJ
Frontiers in immunology 2018;9:945
Frontiers in immunology 2018;9:945
Malaria-induced NLRP12/NLRP3-dependent caspase-1 activation mediates inflammation and hypersensitivity to bacterial superinfection.
Ataide MA, Andrade WA, Zamboni DS, Wang D, Souza Mdo C, Franklin BS, Elian S, Martins FS, Pereira D, Reed G, Fitzgerald KA, Golenbock DT, Gazzinelli RT
PLoS pathogens 2014 Jan;10(1):e1003885
PLoS pathogens 2014 Jan;10(1):e1003885
Frailty in old age is associated with decreased interleukin-12/23 production in response to toll-like receptor ligation.
Compté N, Zouaoui Boudjeltia K, Vanhaeverbeek M, De Breucker S, Tassignon J, Trelcat A, Pepersack T, Goriely S
PloS one 2013;8(6):e65325
PloS one 2013;8(6):e65325
CD56-positive haematological neoplasms of the skin: a multicentre study of the Cutaneous Lymphoma Project Group of the European Organisation for Research and Treatment of Cancer.
Assaf C, Gellrich S, Whittaker S, Robson A, Cerroni L, Massone C, Kerl H, Rose C, Chott A, Chimenti S, Hallermann C, Petrella T, Wechsler J, Bagot M, Hummel M, Bullani-Kerl K, Bekkenk MW, Kempf W, Meijer CJ, Willemze R, Sterry W
Journal of clinical pathology 2007 Sep;60(9):981-9
Journal of clinical pathology 2007 Sep;60(9):981-9
Epithelial cells trigger frontline immunoglobulin class switching through a pathway regulated by the inhibitor SLPI.
Xu W, He B, Chiu A, Chadburn A, Shan M, Buldys M, Ding A, Knowles DM, Santini PA, Cerutti A
Nature immunology 2007 Mar;8(3):294-303
Nature immunology 2007 Mar;8(3):294-303
Local activation of the innate immune system in Buruli ulcer lesions.
Peduzzi E, Groeper C, Schütte D, Zajac P, Rondini S, Mensah-Quainoo E, Spagnoli GC, Pluschke G, Daubenberger CA
The Journal of investigative dermatology 2007 Mar;127(3):638-45
The Journal of investigative dermatology 2007 Mar;127(3):638-45
Expression of the plasmacytoid dendritic cell marker BDCA-2 supports a spectrum of maturation among CD4+ CD56+ hematodermic neoplasms.
Jaye DL, Geigerman CM, Herling M, Eastburn K, Waller EK, Jones D
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2006 Dec;19(12):1555-62
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2006 Dec;19(12):1555-62
TCL1 expression in plasmacytoid dendritic cells (DC2s) and the related CD4+ CD56+ blastic tumors of skin.
Herling M, Teitell MA, Shen RR, Medeiros LJ, Jones D
Blood 2003 Jun 15;101(12):5007-9
Blood 2003 Jun 15;101(12):5007-9
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of normal human peripheral blood cells with Anti-Human CD85g (ILT7) PE (Product # 12-5179-42) and Mouse IgG1 K Isotype Control APC-eFluor® 780 (Product # 47-4714-82) (left) or Anti-Human CD123 APC-eFluor® 780 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 Identification of multiple mDC and pDC subsets by MegaClust. (A) Comparison of the different clusters identified by MegaClust as mDCs and pDCs (example for one patient for four time points). (B) Expression level of CD123 and CD11c of the two mDC clusters and the two pDC clusters identified by MegaClust. (C) Expression levels of HLA-DR and CD4 for the two pDC clusters and the two mDC clusters identified by MegaClust. Cluster 53 could be assigned as basophils due to the lack of expression of CD4 and HLA-DR (). The expression of CD4 is shown for CD4 T cells and CD14 + monocyte clusters for comparison. (D) Overlay of CD4 and HLA-DR of MegaClust identified clusters for mDC, pDC, basophils, CD4 and monocytes. (E) mDCs and pDCs were discriminated according to CD11c and CD123 expression in the supervised flow cytometry re-analysis according to the new gating strategy described in Supplementary Figure 5 (one representative patient 0QZW). (F) Representative illustration of HLA-DR and CD4 co-expression on mDCs and pDCs prior to FACS sorting CD11c+CD123 - CD4 + HLA-DR+ mDCs and CD123 + CD11c - CD4 + HLA-DR + were FACS sorted (dotted lines). (G) Gene expression profiles of flow cytometry sorted CD11c + CD123 - CD4 + HLA-DR + mDCs, CD123 + CD11c - CD4 + HLA-DR + CD4 + HLA-DR + pDC and CD14 + CD16 - monocytes from 3 healthy donors are shown in comparison to the RNA expression profile of public available RNAseq datasets (Immgen) indicating the gene expression profile of B cells, mac
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 1 (A) Surface antigen expression of bone marrow stromal cells (BMSCs) isolated from the talus bone marrow using flow cytometry. >95% of cells were negative for CD45 and CD14, and >99% of cells were negative for CD19 and CD34. In addition, >95% were positive for CD73, CD90, CD105, and CD140b. Since plastic adherence is a well-established and validated technique to isolate multipotent stromal cells (MSCs), we have only characterized one BMSC donor using flowcytometry instead of all donors. The blue lines are stained cells and the gray lines are negative (unstained) controls. Adapted from Croes et al. ( 25 ). (B) Fluorescence-activated cell sorting (FACS) gating strategy used to isolate granulocytes/neutrophils from peripheral blood leukocytes. Granulocytes were either isolated from unlabeled leukocytes using gate 1 (G1) within the forward/sideward scatter (FSC/SSC). Alternatively, leukocytes were stained using CD3, CD14, CD193, and CD123. Within the FSC/SSC of these labeled cells, debris was first excluded [gate 2 (G2)]. Subsequently, CD3+ cells (lymphocytes) and CD14+ (monocytes) were excluded [gate 3 (G3)]. In addition, CD193+ cells (eosinophils) and CD123+ cells (basophils) were excluded [gate 4 (G4)]. The remaining CD3- CD14- CD193- CD123- cells were defined FACS-sorted neutrophils (G2+, G3+, G4+ sorted neutrophils). Re-analysis of FACS-sorted neutrophils shows adequate exclusion of lymphocytes and monocytes based on their FSC/SSC. (C) Images of BMSCs obtained by arr
