- MA1-022 - Invitrogen Antibodies FUT4 antibody

Li T, Zheng Y, Li Y, Ye D
Frontiers in molecular neuroscience 2018;11:392

Ning B, Zhao W, Qian C, Liu P, Li Q, Li W, Wang RF
Nature communications 2017 Aug 24;8(1):349

Ghosh K, Selokar NL, Gahlawat SK, Kumar D, Kumar P, Yadav PS
Animal biotechnology 2016;27(1):38-43

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of SSEA-1 using anti-SSEA-1 monoclonal antibody (Product # MA1-022) shows staining on the cell surface of human differentiated HEL 11.4 iPS cells, indicating loss of pluripotency. SSEA-1 staining (green) and an overlay image of SSEA-1 with DAPI (blue) is shown. HEL 11.4 cells were grown on matrigel coated chamber slides and fixed with formaldehyde prior to staining. Cells were probed with a mouse monoclonal antibody recognizing SSEA-1 (Product # MA1-022) at a dilution of 1:120 over night at 4°C, washed with PBS and incubated with a FITC-conjugated secondary antibody at a dilution of 1:100 for 60 minutes at room temperature. Images were taken at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of SSEA-1 using anti-SSEA-1 monoclonal antibody (Product # MA1-022) shows staining on the cell surface of peripheral human embryonic H9 stem cells, indicating partial loss of pluripotency. SSEA-1 staining (green) and an overlay image of SSEA-1 with DAPI (blue) is shown. H9 cells were grown on matrigel coated chamber slides and fixed with formaldehyde prior to staining. Cells were probed with a mouse monoclonal antibody recognizing SSEA-1 (Product # MA1-022) at a dilution of 1:120 over night at 4°C, washed with PBS and incubated with a FITC-conjugated secondary antibody at a dilution of 1:100 for 60 minutes at room temperature. Images were taken at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of SSEA1 (red) in mouse embryonic CJ7 stem cells grown on 0.1% gelatin. The cells were fixed with 4% paraformaldehyde at room temperature for 10 min and permeabilized with 0.25% Triton-X 100 for 5 min and blocked with 10% BSA in PBS for 30 min at 37°C. Cells were stained with a SSEA1 monoclonal antibody (Product # MA1-022) at a dilution of 1:200 in 3% BSA/PBS blocking buffer overnight at 4°C and then incubated with a RRX-conjugated donkey anti-mouse IgG secondary antibody at a dilution of 1:500 for 1 hour at room temperature. Nucleus DNA (blue) was stained with DAPI (Product # D1306). Note: Data courtesy of Innovators program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of SSEA-1 using anti-SSEA-1 monoclonal antibody (Product # MA1-022) shows specific expression in mouse testicular teratoma F9 cells (shown in green) but not in negative control human NTERA-2 teratocarcinoma cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a mouse monoclonal antibody recognizing SSEA-1 (Product # MA1-022), at a dilution of 1:50 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with a fluorescent dye labeled goat-anti-mouse IgM secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on frozen sections of Knifefish (Apteronotus leptorhynchus) brain tissue fixed with 2% paraformaldehyde and permeabilized with 0.3% Triton X-100. Tissue sections were washed with TBS and blocked in a solution containing 3% serum, 1% BSA, 1% teleostean gelatin, 0.3% Triton X-100 in TBS, for 1 hour at room temperature. The primary antibody SSEA1 (Product # MA1-022X) (1:50 dilution) was applied overnight (~18 hours) at 4°C. After washing with TBS, an Alexa Fluor® 546 - conjugated goat anti-mouse secondary antibody was applied for 2 hours at room temperature at a 1:200 dilution. Sections were counterstained with DAPI. Images were taken with a Zeiss 710 confocal microscope at 63X magnification. The antibody shows specific surface staining of blood vessels in the brain, shown here within a proliferation region in the optic tectum in the brain. In addition, the antibody labeled the surface of certain myelinated axons and did not stain Knifefish stem/progenitor cells. Calibration bar = 10µm. Note: Data courtesy of Innovators program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of SSEA-1 (blue histogram) on H9 human embryonic stem cells. Cells were harvested, fixed with 4% formaldehyde, washed with PBS, and incubated with a SSEA-1 monoclonal antibody (Product # MA1-022) or an IgM isotype control (red histogram) at a 1:100 dilution for 1 hour on ice. For flow analysis, a 30-minute incubation with FITC-conjugated secondary antibody was performed and 100,000 cells were acquired for each sample.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of SSEA-1 (blue histogram) on differentiated HEL 11.4 human iPS cells. Cells were harvested, fixed with 4% formaldehyde, washed with PBS, and incubated with a SSEA-1 monoclonal antibody (Product # MA1-022) or an IgM isotype control (red histogram) at a 1:100 dilution for 1 hour on ice. For flow analysis, a 30-minute incubation with FITC-conjugated secondary antibody was performed and 100,000 cells were acquired for each sample.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1 Derivation and characterization of mouse EpiSCs from in vivo fertilization and NT embryos. (A) Mouse embryo at E5.5 stage, isolated epiblast layer, and morphology of established F-mEpiSCs and NT-mEpiSCs. (B) Immunofluorescence staining for Nanog, SSEA1, and Oct4 in F-EpiSCs and NT-EpiSCs; nuclei are shown using DAPI. (C) Gene expression analysis by qPCR for EpiSC-specific markers, pluripotency markers, and neural marker in four selected mEpiSCs. Scale bar: 100 mum; n.s., P > 0.05; ** P < 0.01.

MA1-022