iPSelector Anti-LNFP I, Human, Mouse-Mono(R-17F) (100 µl)

Product#: FDV-0014B
$400.00
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iPSelector <Anti-LNFP I, Human, Mouse-Mono(R-17F)>(100 µl)

Cat. No. FDV-0014B

Storage -20°C (Avoid repeated freeze-thaw cycles.) 
Volume 100 µL (FDV-0014B), 25 µL (FDV-0014A)
Lot Number see vial label
Host Species and Clonality: Mouse Monoclonal
Clone R-17F
Specificity

This antibody recognizes lacto-N-fucopentaose I (LNFP I: Fucβ1–2Galβ1–3GlcNAcβ1–3Galβ1–4Glc) on a glycolipid / glycoprotein. R-17F epitopes are expressed on undifferentiated human induced pluripotent stem (iPS) / embryonic stem (ES) cells but not on human embryonal carcinoma (EC) cells nor on differentiated human iPS/ES cells.

Isotype and Subclass IgG1
Formulation Phosphate Buffered Saline (PBS) containing 50% Glycerol, contains no preservative.
Purification Protein A Purified
Concentration 1.0 mg/ml
Verified Species Reactivity Human --- Other species not tested.
Immunogen Human iPS cell line, Tic, derived from human fetus lung cells (MRC-5).
Applications 

Western Blot (1:2,000), Immunocytochemistry, Flow Cytometry, Functional Application (Optimal working dilutions should be determined experimentally by each laboratory for each application.)

Antigen- Human 
Host Animal- Mouse 
Cross Reactivity- Human 
Application- FCM, Functional Analysis, IC, IF, Western Blot 
Unlabeled
Purification- PAPu
Class- IgG

Description

New specific antibody for human iPS/ES cells iPSelector (Anti-Human LNFP I, Mouse Monoclonal)
iPSelector is a mouse monoclonal antibody recognizes for lacto-N-fucopentaose I (LNFP I) raised against human iPS cells.
LNFP I is the sugar chain expressed in undifferentiate state human iPS or ES cells then this antibody can be used as a marker for undiffrentiated human iPS or ES cells.

Moreover, this antibody has a function for cytotoxic effect by binding to undifferentiated human iPS or ES cells.
Therefore this antibody is beneficial for removing undifferentiated human iPS or ES cells.

This antibody has been commercialized under a license from Ritsumeikan University.

Features

  • Specific for undifferentiated human iPS or ES cells.
  • Uniformly stain cell membrane of human iPS or ES cells.
  • Can be used for removing undifferentiated human iPS or ES cells by cytotoxic effect.
  • Clone : R-17F
Data
(1) Western Blot
FDV-0014_fig1.png One major positive band and several minor bands were specific to human iPS cells, and any positive band were not obtained with HEK293 cells.

(Condition)
Sample: 5 μg cell lysate in each lane.
Left: human iPS cells (LNFP I positive)
Right: HEK293 (Negative Control)
Dilution: 1:2,000
Secondary antibody: Anti-Mouse IgG, Goat-Poly, HRP 
(Kirkegaard & Perry Laboratories, #074-1516)
Chemiluminescence Substrate: Trident plus Western HRP Substrate 
(GeneTex International Corporation, #GTX400006)
Detection: LuminoGraph I (ATTO) with 1min exposure


(2) Immunocytochemical Staining

FDV-0014_fig2.png
Cultured human iPS cells were stained with R-17F, SSEA-3, and SSEA-4 antibodies. [bars: 100 μm]
R-17F stained the entire surface of the cell membranes equally, while the staining by SSEA-3 and SSEA-4 antibodies are not evenly. 
This suggests that R-17F epitope is expressed ubiquitously all over the human iPS cells.

(3) Functional Application (Cytotoxic effects on undifferentiated iPS/ES cells)

FDV-0014_fig3.png 
R-17F is reported to exhibit potent dose-dependent cytotoxicity when it is added to living human iPS/ES cells. 
[Left] After the incubation of iPS cell suspension with R-17F at 4°C for only 45 minutes, the percentage of viable cells decreased concentration-dependently (red triangles). 
Blue squares: effects of the isotype (IgG1)-matching control antibody (anti-α-MBP) as Negative Control
[Right] When R-17F-treated iPS cells were incubated with a small amount (0.025-0.1 μg) of the secondary antibody (goat anti-mouse IgG1 antibody), the cytotoxic effect of R-17F was enhanced significantly in a dose-dependent manner (red triangles).

Comparison table : Binding Activity of Antibodies to Cells

  R-17F TRA-1-60 TRA-1-81 SSEA-3 SSEA-4
Tic (iPS) ++++ ++++ ++++ ++++ ++++
KhES-3 (ES) +++ ++++ ++++ +++ ++++
H9 (ES) ++++ ++++ ++++ +++ ++++
2102Ep (EC) +/- ++++ ++++ +++ +++

iPSelector (Clone : R-17F)

Product Background

Clone SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81 antibodies are well-known as human iPS/ES cell-marker antibodies. Since SSEA-3 antibody was originally generated against mouse embryo and the other antibodies were against human EC cells, these antibodies recognize not only human iPS/ES cells but also human EC cells.

R-17F is a novel mouse monoclonal antibody generated by using a human iPS cell line as an immunogen. It is specific to human iPS/ES cells and does not essentially cross-react against human EC cells (Table, ref. 1). 
This R-17F antibody also stains entire surface of human iPS/ES cell membranes evenly, while the staining by SSEA-3 and SSEA-4 antibodies are not uniformly (ref. 2). 

In addition, R-17F is reported to exhibit potent dose-dependent cytotoxicity when added to living human iPS/ES cells (ref. 2 & 3).
R-17F is a beneficial tool for the selective detection, staining and removal of undifferentiated of human iPS/ES cells in regenerative medicine.

References

  1. Kawabe, K., et al., Glycobiology, 23, 322, (2013).
  2. Matsumoto, S., et al., J. Biol. Chem., 290, 20071, (2015).
  3. Nakao, H., et al., Glycoconj. J., DOI 10.1007/s10719-0169-9710-2
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