Detection of Human EOMES by Western Blot.
Western blot shows lysates of BG01V human embryonic stem cells untreated (-) or mesoendoderm differentiated (+). PVDF Membrane was probed with 1 µg/mL of Human EOMES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6166) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for EOMES at approximately 85-105 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
EOMES in mesoderm lineage cells differentiated from BG01V.
EOMES was detected in immersion fixed BG01V human embryonic stem cells differentiated into mesoderm using Human EOMES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6166) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of EOMES in Differentiated BG01V Human Cells by Flow Cytometry.
BG01V human embryonic stem cells differentiated to mesendoderm were stained with Sheep Anti-Human EOMES Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6166, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
EOMES (Eomesodermin; Eo from Greek meaning "dawn"/early in mesoderm; also TBR2) is a 72 kDa member of the TBR1 subfamily, T-box family of transcription factors. It is expressed in NK and CD8+ T cells, where CTLA4 activation suppresses EOMES activation of IFN-gamma and granzyme B genes. It is also found in the embryo, where it occurs in forebrain floorplate and migrating neuroblasts at 12.5 weeks gestation. Notably, it is reported to undergo intercellular transfer in fetal Xenopus tissue destined to become mesoderm. Here, it synchronizes a multicellular commitment to a cell lineage. Human EOMES is 686 amino acids (aa) in length. It contains short poly-Ala, -Gly and -Asn motifs, and a DNA-binding T box (aa 276-456). There is one isoform that shows a 19 aa insertion after Ser460. Over aa 471‑686, human EOMES shares 91% aa identity with mouse EOMES.
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