SSEA4 Antibody [E20H6] | Primary Antibodies

Application: Reactivity: Human

Usage Information

Dilution
IF1:100 - 1:400 FCM1:200 - 1:800

Application
IF, FCM

Reactivity
Human

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Positive Control	NTERA-2 cells
Negative Control	Jurkat cells

Datasheet & SDS

Biological Description

Specificity
SSEA4 Antibody [E20H6] detects endogenous levels of total SSEA4 protein.

Clone
E20H6

Background
SSEA4 (Stage-Specific Embryonic Antigen 4) is a sialylated hexasaccharide glycolipid epitope (Neu5Acα2-3Galβ1-3GalNAcβ1-3Galα1-4Galβ1-4Glc-Cer) found on human embryonic stem cells (hESCs), induced pluripotent stem cells (iPSCs), teratocarcinoma cells, and cancer stem cells, but absent from differentiated somatic or mouse pluripotent cells. Its carbohydrate headgroup adopts a horseshoe-shaped conformation recognized by MC813 antibodies through a network of hydrogen bonds involving the terminal sialic acid (Neu5Ac) carboxylate with Asp54H/Gly53H and N-acetyl GalNAc hydroxyls with Asp49L, which underpins its utility for specific detection of pluripotency states via flow cytometry and immunohistochemistry. The glycan’s rigid α-Gal linkage and β1-3 glycosidic bonds maintain a stable, extended conformation atop ceramide lipid tails in plasma membranes, where SSEA4 actively supports stemness by modulating Wnt/β-catenin signaling through direct interaction with the LRP6 co-receptor, enabling ligand-independent pathway activation, and by recruiting PI3K/Akt through glycan-mediated clustering that sustains Nanog/Oct4/Sox2 circuitry essential for self-renewal. During differentiation, downregulation of glycosyltransferases (ST3GAL6, B3GALT5) progressively removes the terminal sialic acid, converting SSEA4 to SSEA3 and extinguishing its signaling competence.

Background

SSEA4 (Stage-Specific Embryonic Antigen 4) is a sialylated hexasaccharide glycolipid epitope (Neu5Acα2-3Galβ1-3GalNAcβ1-3Galα1-4Galβ1-4Glc-Cer) found on human embryonic stem cells (hESCs), induced pluripotent stem cells (iPSCs), teratocarcinoma cells, and cancer stem cells, but absent from differentiated somatic or mouse pluripotent cells. Its carbohydrate headgroup adopts a horseshoe-shaped conformation recognized by MC813 antibodies through a network of hydrogen bonds involving the terminal sialic acid (Neu5Ac) carboxylate with Asp54H/Gly53H and N-acetyl GalNAc hydroxyls with Asp49L, which underpins its utility for specific detection of pluripotency states via flow cytometry and immunohistochemistry. The glycan’s rigid α-Gal linkage and β1-3 glycosidic bonds maintain a stable, extended conformation atop ceramide lipid tails in plasma membranes, where SSEA4 actively supports stemness by modulating Wnt/β-catenin signaling through direct interaction with the LRP6 co-receptor, enabling ligand-independent pathway activation, and by recruiting PI3K/Akt through glycan-mediated clustering that sustains Nanog/Oct4/Sox2 circuitry essential for self-renewal. During differentiation, downregulation of glycosyltransferases (ST3GAL6, B3GALT5) progressively removes the terminal sialic acid, converting SSEA4 to SSEA3 and extinguishing its signaling competence.

References
https://pubmed.ncbi.nlm.nih.gov/6141938/ https://pubmed.ncbi.nlm.nih.gov/31831622/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%