Anti-Mouse SSEA-1 (CD15) Antibody, Clone MC-480

Mouse monoclonal IgM antibody against human, mouse, rat SSEA-1 (CD15)
概要
The MC-480 antibody reacts with a terminal carbohydrate epitope, stage-specific embryonic antigen-1 (SSEA-1), which is expressed on a large-molecular-mass (> 200 kDa) glycoprotein on the surface of early mouse embryos, mouse embryonal carcinoma (EC), embryonic stem (ES) cells and mouse and human embryonic germ (EG) cells. SSEA-1 is not expressed on undifferentiated human EC, ES or induced pluripotent stem (iPS) cells, or rhesus monkey ES cell lines. Its expression on mouse ES cells is decreased upon differentiation, whereas in humans, expression is upregulated during differentiation. SSEA-1 is also found on adult human granulocytes and monocytes, where it is denoted CD15, and the MC-480 antibody recognizes the CD15 marker on these cell types. It has been reported that SSEA-1 has roles in cell adhesion and migration, and regulation of cell differentiation.

This antibody clone has been verified for purity assessments of cells isolated with EasySep™ kits, including EasySep™ HLA Whole Blood CD15 Positive Selection Kit (Catalog #18681HLA; partial blocking may be observed), and for labeling human ES and iPS cells grown in TeSR™-E8™ (Catalog #05940), mTeSR™1 (Catalog #85850) and TeSR™2 (Catalog #05860).
Subtype
Primary Antibodies
Target Antigen
SSEA-1 (CD15)
Alternative Names
3-FAL, CD15, Lewis X, SSEA1, Stage-specific embryonic antigen 1, X-hapten
Reactive Species
Human, Mouse, Rat
Conjugation
Alexa Fluor 488, Biotin, FITC, PE, Unconjugated
Host Species
Mouse
Cell Type
Pluripotent Stem Cells
Species
Human, Mouse, Rat
Application
Cell Isolation, Flow Cytometry, Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blotting
Area of Interest
Stem Cell Biology
Clone
MC-480
Gene ID
14345
Isotype
IgM, kappa
数据及文献

Data

Data for Alexa Fluor® 488-Conjugated

Figure 1. Data for Alexa Fluor® 488-Conjugated

(A) Flow cytometry analysis of human whole blood nucleated cells labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, Alexa Fluor® 488 (filled histogram) or Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, Alexa Fluor® 488 (Catalog #60069AD) (solid line histogram). SSEA-1 is highly expressed on granulocytes.
(B) Flow cytometry analysis of human HT1080 fibrosarcoma cells labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, Alexa Fluor® 488 (filled histogram). Labeling of human HT1080 fibrosarcoma (solid line histogram) or ES cells (negative control; dashed line histogram) with a mouse IgM, kappa isotype control antibody (Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R, Alexa Fluor® 488; Catalog #60064AD) is shown. SSEA-1 is not expressed on undifferentiated human ES cells.
(C) Human ES cells were cultured in mTeSR™1 on Corning® Matrigel®-coated glass slides, then fixed and labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, Alexa Fluor® 488. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, Alexa Fluor® 488.

Data for Biotin-Conjugated

Figure 2. Data for Biotin-Conjugated

(A) Flow cytometry analysis of human buffy coat nucleated cells labeled with Anti-Human SSEA-1 Antibody, Clone MC-480, Biotin, followed by streptavidin (SAV) APC (filled histogram), or a mouse IgM, kappa biotin isotype control antibody, followed by SAV APC (solid line histogram). (B) Flow cytometry analysis of human buffy coat nucleated cells processed with the EasySep™ HLA Whole Blood CD15 Positive Selection Kit and labeled with Anti-Human SSEA-1 Antibody, Clone MC-480, Biotin, followed by SAV APC. Histograms show labeling of buffy coat nucleated cells (Start) and isolated cells (Isolated). Labeling with a mouse IgM, kappa biotin isotype control antibody, followed by SAV APC is shown (solid line histogram).

Data for FITC-Conjugated

Figure 3. Data for FITC-Conjugated

(A) Flow cytometry analysis of human buffy coat nucleated cells labeled with Anti-Human SSEA-1 Antibody, Clone MC-480, FITC (filled histogram) or a mouse IgM, kappa FITC Isotype control antibody (solid line histogram). SSEA-1 (CD15) is highly expressed on granulocytes. (B) Flow cytometry analysis of human buffy coat nucleated cells processed with the EasySep™ HLA CD15 WB Positive Selection Kit and labeled with Anti-Human SSEA-1 Antibody, Clone MC-480, FITC. Histograms show labeling of buffy coat nucleated cells (Start) and isolated cells (Isolated). Labeling of start cells with a mouse IgM, kappa FITC Isotype control antibody is shown (solid line histogram).

Data for PE-Conjugated

Figure 4. Data for PE-Conjugated

(A) Flow cytometry analysis of human whole blood nucleated cells labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, PE (filled histogram) or Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE (Catalog #60069PE) (solid line histogram). SSEA-1 is highly expressed on granulocytes.
(B) Human ES cells were cultured in mTeSR™1 on Corning® Matrigel®-coated glass slides, then fixed and labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, PE. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE. SSEA-1 is not expressed on undifferentiated human ES cells.
(C) Flow cytometry analysis of human buffy coat nucleated cells processed with the EasySep™ HLA CD15 WB Positive Selection Kit and labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, PE. Histograms show labeling of buffy coat nucleated cells (Start) and isolated cells (Isolated). Labeling with Mouse IgM, kappa Isotype Control Antibody, Clone MM-30, PE is shown (solid line histogram).

Data for Unconjugated

Figure 5. Data for Unconjugated

(A) Flow cytometry analysis of human whole blood nucleated cells labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, followed by Goat Anti-Mouse IgG (H+L) Antibody, Polyclonal, FITC (Catalog #60138FI) (filled histogram), or Mouse IgM, kappa Isotype Control Antibody, Clone MM-30 (Catalog #60069), followed by Goat Anti-Mouse IgG (H+L) Antibody, Polyclonal, FITC (solid line histogram). SSEA-1 is highly expressed on granulocytes.
(B) Flow cytometry analysis of human HT1080 fibrosarcoma cells labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, followed by goat antimouse IgG, FITC (filled histogram). Labeling of human HT1080 fibrosarcoma cells (solid line histogram) or H1 ES cells (negative control; dashed line histogram) with a mouse IgM, kappa isotype control antibody (Anti-Human TRA-1-60 Antibody, Clone TRA-1-60R; Catalog #60064) is shown.
(C) Human ES cells were cultured in mTeSR™1 on Corning® Matrigel®-coated glass slides, then fixed and labeled with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480, followed by goat anti-mouse IgG, FITC. Inset shows cells labeled with Mouse IgM, kappa Isotype Control Antibody, Clone MM- 30, followed by goat anti-mouse IgG, FITC. SSEA-1 is not expressed on undifferentiated human ES cells.
(D) Western blot analysis of denatured/reduced cell lysates from human ES cells (negative control; lane 1) or HT1080 fibrosarcoma cells (lane 2) with Anti-Human SSEA-1 (CD15) Antibody, Clone MC-480.

Publications (1)

Cell death & disease 2014 JAN OCT4 as a target of miR-34a stimulates p63 but inhibits p53 to promote human cell transformation Ng WL et al.
Abstract
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Abstract

Human cell transformation is a key step for oncogenic development, which involves multiple pathways; however, the mechanism remains unclear. To test our hypothesis whether cell oncogenic transformation shares some mechanisms with the process of reprogramming non-stem cells to induced pluripotent stem cells (iPSC), we studied the relationship among the key factors for promoting or inhibiting iPSC in radiation-transformed human epithelial cell lines derived from different tissues (lung, breast and colon). We unexpectedly found that p63 and OCT4 were highly expressed (accompanied by low expressed p53 and miR-34a) in all transformed cell lines examined when compared with their non-transformed counterparts. We further elucidated the relationship of these factors: the 3p strand of miR-34a directly targeted OCT4 by binding to the 3′ untranslated region (3′-UTR) of OCT4 and, OCT4, in turn, stimulated p63 but inhibited p53 expression by binding to a specific region of the p63 or p53 promoter. Moreover, we revealed that the effects of OCT4 on promoting cell oncogenic transformation were by affecting p63 and p53. These results support that a positive loop exists in human cells: OCT4 upregulation as a consequence of inhibition of miR-34a, promotes p63 but suppresses p53 expression, which further stimulates OCT4 upregulation by downregulating miR-34a. This functional loop contributes significantly to cell transformation and, most likely, also to the iPSC process.
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