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iPS/ES cells specific monoclonal antibody Anti-Lacto-N-fucopentaose I (LNFP I), Mouse-Mono (R-17F)

Date:November 06 2017Web Page No:80958

Funakoshi Co.,Ltd.

New specific antibody for human iPS/ES cells
Anti-Lacto-N-fucopentaose I (LNFP I), Mouse-Mono (R-17F)

Anti-LNFP I 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

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Data

Western Blot

Verification by WB

5 ug cell lysate in each lane.

  • Left: human iPS cells (LNFP I positive)
  • Right: HEK293 (LNFP I negative)
  • Dilution: 1:2,000 (0.5 μg/ml)
  • Secondary antibody: Anti-Mouse IgG, Goat-Poly, HRP (Kirkegaard & Perry Laboratories, #5220-0337)
  • Chemiluminescence Substrate: Trident plus Western HRP Substrate (GeneTex, #GTX400006)
  • Detection: LuminoGraph I (ATTO) with 1min exposure

One major positive band and several minor bands were specific to human iPS cells, and any positive band was not obtained with HEK293 cells.


Immunocytochemical Staining

IHC

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.


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

R-17F is reported to exhibit potent dose-dependent cytotoxicity when it is added to living human iPS/ES cells.

Confirmation of cytotoxic activity
  • [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).

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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) +/- ++++ ++++ +++ +++

* Anti-LNFP I (Clone : R-17F)

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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.


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Reference

  1. Kawabe, K., et al., "A novel antibody for human induced pluripotent stem cells and embryonic stem cells recognizes a type of keratan sulfate lacking oversulfated structures", Glycobiology, 23(3), 322~336, (2013). [PMID:23154990]
  2. Matsumoto, S., et al., "A Cytotoxic Antibody Recognizing Lacto-N-fucopentaose I (LNFP I) on Human Induced Pluripotent Stem (hiPS) Cells", J. Biol. Chem., 290(33), 20071~20085, (2015) [PMID:26100630].
  3. Nakao, H., et al., "Characterization of glycoproteins expressing the blood group H type 1 epitope on human induced pluripotent stem (hiPS) cells", Glycoconj. J., 34, 779-787 (2017) [PMID:27431816].

[Date : May 30 2024 00:08]

Detail Product Name Product Code Supplier Size Price
Anti-Lacto-N-fucopentanose I (LNFP I), Mouse-Mono (R-17F)
DatasheetThis may not be the latest data sheet.
FDV-0014B FNAFunakoshi Co.,Ltd. 100 µl $400

Description Anti-LNFP I 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.
Storage -20°C
Antigen Species Host Species Mouse Class IgG Label Unlabeled
Cross Reactivity Human Application FCM,Functional Analysis,IC,IF,Western Blot
Clonality Format Purification Protein A/G Affinity Purified Absorption
Link

[Date : May 30 2024 00:08]

Anti-Lacto-N-fucopentanose I (LNFP I), Mouse-Mono (R-17F)


  • Product Code: FDV-0014B
  • Supplier: FNA
  • Size: 100µl
  • Price: $400

Description Anti-LNFP I 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.
Storage -20°C
Antigen Species Host Species Mouse
Class IgG Label Unlabeled
Cross Reactivity Human Application FCM,Functional Analysis,IC,IF,Western Blot
Clonality Format
Purification Protein A/G Affinity Purified Absorption
Link

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