CD133/2 Antibody, anti-human, APC

Protocol tips

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The cultured glioma cells were digested by trypsin or accutase and resuspended with PBS. The fresh glioma specimens were transferred to laboratory on ice in half hour after surgery, then washed and enzymatically dissociated into single cells and resuspended in PBS		The FACS analysis and cell sorting were performed on BD FACS Aria II cytometer (USA) or Beckman moflo XDP (USA).

Upstream tips
The cultured glioma cells were digested by trypsin or accutase and resuspended with PBS. The fresh glioma specimens were transferred to laboratory on ice in half hour after surgery, then washed and enzymatically dissociated into single cells and resuspended in PBS
Downstream tips
The FACS analysis and cell sorting were performed on BD FACS Aria II cytometer (USA) or Beckman moflo XDP (USA).

Publication protocol

"The cultured glioma cells were digested by trypsin or accutase and resuspended with PBS. The fresh glioma specimens were transferred to laboratory on ice in half hour after surgery, then washed and enzymatically dissociated into single cells and resuspended in PBS. The staining procedures for CD133 and CD15 markers were performed as previously described [6, 8]. The labeling antibodies were anti-CD133-APC antibody (Clone REA816; Miltenyi Biotec, Germany) and anti-CD15-FITC antibody (Biolegend, USA) with REA Control (S)-APC (Miltenyi Biotec, Germany) and FITC Mouse IgM (Biolegend, USA) as controls, respectively.

The FACS analysis and cell sorting were performed on BD FACS Aria II cytometer (USA) or Beckman moflo XDP (USA). For analyzing and sorting with NADH autofluorescence intensity as a marker, an excitation wavelength of 375 nm or 355 nm and an emission wavelength of 450/50BP filter were used. For analyzing and sorting with CD133 and CD15 as markers, labeled cells were analyzed and sorted with corresponding excitation and emission wavelengths of the fluorochrome. All data were analyzed with BD FACSDiva software version 8 or Beckman moflo XDP submmit 5.2."

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