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The sequential gating strategy was used to identify immune cell populations from healthy human samples. Healthy donor samples, either ( A ) cryopreserved peripheral blood mononuclear cells (cPBMCs) or ( B ) fresh whole blood (WB), were stimulated for 15 min, fixed, palladium-barcoded, and stained with a comprehensive panel of 19 (for cPBMCs) or 20 (for whole blood) surface markers. Within the single-cell gate, non-granulocytes (leukocytes) were defined as CD66b-CD45 + , while granulocytes were confirmed solely in whole blood as CD66b + CD45-. Lymphocytes were resolved using CD3 and CD56 [T cells (CD3 + CD56-)], NKT cells (CD3 + CD56 + ), and NK cells (CD3-CD56 + ). B cells were identified as CD19 + CD20 +/- within the CD3-CD56- gate, further confirmed by negative expression of <t>CD123</t> and CD11c. Both T and B cells were further subset based on the expression of CD4, CD8, CD45RA, CD27, and CD25 or IgD and CD27, respectively. Non-T, non-B, and non-NK cells (CD45 + CD3-CD19-CD56-CD20-) were separated based on their expression of CD11c and HLA-DR. Total DCs were defined as CD11c + HLA-DR + and can be further defined through the expression of CD123 (plasmacytoid DCs). Monocytes were resolved within this non-lymphocyte gate based on their expression of CD14 and CD16.
Cd123 Il 3r, supplied by fluidigm, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fluidigm 3151001b
The sequential gating strategy was used to identify immune cell populations from healthy human samples. Healthy donor samples, either ( A ) cryopreserved peripheral blood mononuclear cells (cPBMCs) or ( B ) fresh whole blood (WB), were stimulated for 15 min, fixed, palladium-barcoded, and stained with a comprehensive panel of 19 (for cPBMCs) or 20 (for whole blood) surface markers. Within the single-cell gate, non-granulocytes (leukocytes) were defined as CD66b-CD45 + , while granulocytes were confirmed solely in whole blood as CD66b + CD45-. Lymphocytes were resolved using CD3 and CD56 [T cells (CD3 + CD56-)], NKT cells (CD3 + CD56 + ), and NK cells (CD3-CD56 + ). B cells were identified as CD19 + CD20 +/- within the CD3-CD56- gate, further confirmed by negative expression of <t>CD123</t> and CD11c. Both T and B cells were further subset based on the expression of CD4, CD8, CD45RA, CD27, and CD25 or IgD and CD27, respectively. Non-T, non-B, and non-NK cells (CD45 + CD3-CD19-CD56-CD20-) were separated based on their expression of CD11c and HLA-DR. Total DCs were defined as CD11c + HLA-DR + and can be further defined through the expression of CD123 (plasmacytoid DCs). Monocytes were resolved within this non-lymphocyte gate based on their expression of CD14 and CD16.
3151001b, supplied by fluidigm, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ch dvs fluidigm 3089003b 467 cd56 163dy cd56 163dy false ncam16 2 hu dvs fluidigm 3163007b 536 cd123  (fluidigm)
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fluidigm ch dvs fluidigm 3089003b 467 cd56 163dy cd56 163dy false ncam16 2 hu dvs fluidigm 3163007b 536 cd123
The sequential gating strategy was used to identify immune cell populations from healthy human samples. Healthy donor samples, either ( A ) cryopreserved peripheral blood mononuclear cells (cPBMCs) or ( B ) fresh whole blood (WB), were stimulated for 15 min, fixed, palladium-barcoded, and stained with a comprehensive panel of 19 (for cPBMCs) or 20 (for whole blood) surface markers. Within the single-cell gate, non-granulocytes (leukocytes) were defined as CD66b-CD45 + , while granulocytes were confirmed solely in whole blood as CD66b + CD45-. Lymphocytes were resolved using CD3 and CD56 [T cells (CD3 + CD56-)], NKT cells (CD3 + CD56 + ), and NK cells (CD3-CD56 + ). B cells were identified as CD19 + CD20 +/- within the CD3-CD56- gate, further confirmed by negative expression of <t>CD123</t> and CD11c. Both T and B cells were further subset based on the expression of CD4, CD8, CD45RA, CD27, and CD25 or IgD and CD27, respectively. Non-T, non-B, and non-NK cells (CD45 + CD3-CD19-CD56-CD20-) were separated based on their expression of CD11c and HLA-DR. Total DCs were defined as CD11c + HLA-DR + and can be further defined through the expression of CD123 (plasmacytoid DCs). Monocytes were resolved within this non-lymphocyte gate based on their expression of CD14 and CD16.
Ch Dvs Fluidigm 3089003b 467 Cd56 163dy Cd56 163dy False Ncam16 2 Hu Dvs Fluidigm 3163007b 536 Cd123, supplied by fluidigm, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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IL-3 boosts BCSC marker expression and mammosphere formation. ( A , B ) Representative FACS dot plots of ALDH1A1 and CD44/CD24 in MDA-MB-436 and ( C , D ) Hs-578T cells upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( E , F ) ALDH activity assay on supernatant of MDA-MB-436 and Hs-578T cell culture upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( G-H ) Mammosphere formation assay of MDA-MB-436 and Hs-578T cells subjected to treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5ng/ml), NC (100 pmol) + IL-3 (5 ng/ml), <t>IL-3Rα</t> blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) and IL-3 blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) (n = 3; 3 biological replicates for each n). Original magnification corresponds to 20X, scale bar: 50 µm. Data are represented as mean ± SEM
Il 3rα Apc, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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IL-3 boosts BCSC marker expression and mammosphere formation. ( A , B ) Representative FACS dot plots of ALDH1A1 and CD44/CD24 in MDA-MB-436 and ( C , D ) Hs-578T cells upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( E , F ) ALDH activity assay on supernatant of MDA-MB-436 and Hs-578T cell culture upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( G-H ) Mammosphere formation assay of MDA-MB-436 and Hs-578T cells subjected to treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5ng/ml), NC (100 pmol) + IL-3 (5 ng/ml), <t>IL-3Rα</t> blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) and IL-3 blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) (n = 3; 3 biological replicates for each n). Original magnification corresponds to 20X, scale bar: 50 µm. Data are represented as mean ± SEM
Cd123, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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The sequential gating strategy was used to identify immune cell populations from healthy human samples. Healthy donor samples, either ( A ) cryopreserved peripheral blood mononuclear cells (cPBMCs) or ( B ) fresh whole blood (WB), were stimulated for 15 min, fixed, palladium-barcoded, and stained with a comprehensive panel of 19 (for cPBMCs) or 20 (for whole blood) surface markers. Within the single-cell gate, non-granulocytes (leukocytes) were defined as CD66b-CD45 + , while granulocytes were confirmed solely in whole blood as CD66b + CD45-. Lymphocytes were resolved using CD3 and CD56 [T cells (CD3 + CD56-)], NKT cells (CD3 + CD56 + ), and NK cells (CD3-CD56 + ). B cells were identified as CD19 + CD20 +/- within the CD3-CD56- gate, further confirmed by negative expression of CD123 and CD11c. Both T and B cells were further subset based on the expression of CD4, CD8, CD45RA, CD27, and CD25 or IgD and CD27, respectively. Non-T, non-B, and non-NK cells (CD45 + CD3-CD19-CD56-CD20-) were separated based on their expression of CD11c and HLA-DR. Total DCs were defined as CD11c + HLA-DR + and can be further defined through the expression of CD123 (plasmacytoid DCs). Monocytes were resolved within this non-lymphocyte gate based on their expression of CD14 and CD16.

Journal: Bio-protocol

Article Title: Dual Phospho-CyTOF Workflows for Comparative JAK/STAT Signaling Analysis in Human Cryopreserved PBMCs and Whole Blood

doi: 10.21769/BioProtoc.5512

Figure Lengend Snippet: The sequential gating strategy was used to identify immune cell populations from healthy human samples. Healthy donor samples, either ( A ) cryopreserved peripheral blood mononuclear cells (cPBMCs) or ( B ) fresh whole blood (WB), were stimulated for 15 min, fixed, palladium-barcoded, and stained with a comprehensive panel of 19 (for cPBMCs) or 20 (for whole blood) surface markers. Within the single-cell gate, non-granulocytes (leukocytes) were defined as CD66b-CD45 + , while granulocytes were confirmed solely in whole blood as CD66b + CD45-. Lymphocytes were resolved using CD3 and CD56 [T cells (CD3 + CD56-)], NKT cells (CD3 + CD56 + ), and NK cells (CD3-CD56 + ). B cells were identified as CD19 + CD20 +/- within the CD3-CD56- gate, further confirmed by negative expression of CD123 and CD11c. Both T and B cells were further subset based on the expression of CD4, CD8, CD45RA, CD27, and CD25 or IgD and CD27, respectively. Non-T, non-B, and non-NK cells (CD45 + CD3-CD19-CD56-CD20-) were separated based on their expression of CD11c and HLA-DR. Total DCs were defined as CD11c + HLA-DR + and can be further defined through the expression of CD123 (plasmacytoid DCs). Monocytes were resolved within this non-lymphocyte gate based on their expression of CD14 and CD16.

Article Snippet: 151 Eu , CD123 (IL-3R) , Standard BioTools , 3151001B , 6H6 , 8.75.

Techniques: Staining, Expressing

IL-3 boosts BCSC marker expression and mammosphere formation. ( A , B ) Representative FACS dot plots of ALDH1A1 and CD44/CD24 in MDA-MB-436 and ( C , D ) Hs-578T cells upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( E , F ) ALDH activity assay on supernatant of MDA-MB-436 and Hs-578T cell culture upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( G-H ) Mammosphere formation assay of MDA-MB-436 and Hs-578T cells subjected to treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5ng/ml), NC (100 pmol) + IL-3 (5 ng/ml), IL-3Rα blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) and IL-3 blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) (n = 3; 3 biological replicates for each n). Original magnification corresponds to 20X, scale bar: 50 µm. Data are represented as mean ± SEM

Journal: Breast Cancer Research : BCR

Article Title: IL-3/STAT5/miR-155-5p axis supports stem-related pathway reprogramming in TNBC

doi: 10.1186/s13058-025-02143-1

Figure Lengend Snippet: IL-3 boosts BCSC marker expression and mammosphere formation. ( A , B ) Representative FACS dot plots of ALDH1A1 and CD44/CD24 in MDA-MB-436 and ( C , D ) Hs-578T cells upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( E , F ) ALDH activity assay on supernatant of MDA-MB-436 and Hs-578T cell culture upon treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5 ng/ml) and NC (100 pmol) + IL-3 (5 ng/ml) for 24h (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM. ( G-H ) Mammosphere formation assay of MDA-MB-436 and Hs-578T cells subjected to treatment with saline, IL-3 (5 ng/ml), antagomiR-155 (100 pmol) + IL-3 (5ng/ml), NC (100 pmol) + IL-3 (5 ng/ml), IL-3Rα blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) and IL-3 blocking mAb (1 ng/ml) + IL-3 (5 ng/ml) (n = 3; 3 biological replicates for each n). Original magnification corresponds to 20X, scale bar: 50 µm. Data are represented as mean ± SEM

Article Snippet: The following anti-human antibodies were used: IL-3Rα-APC, (Milteny Biotec #130-113-322), CD44-FITC (Miteny Biotec #130-113-341), CD24-PE (Milteny Biotec #130-112-656), ALDH1A1-PE (abcam #ab209537).

Techniques: Marker, Expressing, Saline, Activity Assay, Cell Culture, Tube Formation Assay, Blocking Assay