Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 64-1239-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD123 Monoclonal Antibody (6H6), Super Bright™ 645, 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-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.5 µg/mL) 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. Super Bright 645 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 645 nm. We recommend using a 660/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. 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: 405 nm; Emission: 645 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 6H6
- Vial size
- 100 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.
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
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells were stained with CD85g Monoclonal Antibody, APC (Product # 17-5179-42) and Mouse IgG1 kappa Isotype Control, Super Bright 645 (Product # 64-4714-82) (left) or CD123 Monoclonal Antibody, Super Bright 645 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
- 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