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rBC2LCN Stripping Solution

rBC2LCN (AiLecS1) is a recombinant lectin which has high specificity to sugar chains which exists on the cell surfaces of undifferentiated human ES and human iPS cells(hPSCs). Therefore it is used as a marker ofundifferentiated hPSCs.
This product strips away rBC2LCN lectin bound to sugar chains on the surfaces of undifferentiated hPSCs. After the lectin is removed, hPSCs can be stained with other antibodies or continued to be cultured.

Features

  • Can strip rBC2LCN from cells with treatment for 10 – 30 minutes
  • Can be used straight from the container (no formulation required)
  • Low cytotoxicity
  • Can be used after both live cell imaging and fixed cell staining

Product outline

  • Sterility test passed (filtration sterilization performed with a 0.1 μm filter)
  • Composition: Hanks' Balanced Salt Solution Modified

Instructions for use

  1. Prepare cells stained with rBC2LCN
  2. Aspirate the culture media and add rBC2LCN Stripping Solution
  3. Incubate for 10-30 minutes at 37°C with 5% CO2
  4. Aspirate the product and add new culture media (add D-PBS or any other suitable solution if staining will be repeated with another antibody)
  5. Continue culture or stain with other antibodies (*In the case of live cell imaging, avoid incubation with the rBC2LCN Stripping Solution lasting more than 30 minutes)

Removal of rBC2LCN-FITC bound to the surface of human iPS cells

rBC2LCN-FITC was added at a ratio of 1/100 to culture media containing human iPS cell 201B7 strain. After 35 minutes of staining, the culture media was aspirated, rBC2LCN Stripping Solution was added and incubated for 30 minutes. Flow cytometry was then performed on cells treated with rBC2LCN-FITC and with rBC2LCN Stripping Solution

Colony staining image

Human iPS cells were stained with rBC2LCN-FITC. Thereafter, by adding rBC2LCN Stripping Solution, rBC2LCN-FITC was stripped from cell surface, and no fluorescence was observed on the cell surface..

00573_img01.jpg

Flow cytometry

Human iPS cell 201B7 strain stained with rBC2LCN-FITC was treated with rBC2LCN Stripping Solution. Flow cytometry revealed a shift to the left of the cell count peak for treated cells compared to that for untreated cells.

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Effects of rBC2LCN Stripping Solution on human iPS cells

Human iPS cell 201B7 strain was stained with rBC2LCN-FITC. rBC2LCN-FITC was then stripped by adding rBC2LCN Stripping Solution. Both cells stained with rBC2LCN-FITC and cells treated with rBC2LCN Stripping Solution after rBC2LCN-FITC staining were cultured.
It was consequently found that iPS cells treated with rBC2LCN Stripping Solution displayed the same cell growth properties as untreated controls, confirming that the solution had no effect on cell growth.

00573_img03.jpg

〔Cell〕
human iPS cell 201B7 strain
〔Culture medium composition〕
StemSure® hPSC MediumΔ + 35ng/ml bFGF
〔Coating〕
Matrigel® hESC-Qualified Matrix
〔Seeding density〕
4×104 cells/well(with 12well)
〔Incubation time〕
5 days

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rBC2LCN lectin

StemSure®hPSC MediumΔ

StemSure® series

Product content may differ from the actual image due to minor specification changes etc.

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