Flow cytometry Anti-bodies Human - CD133

Flow cytometry is an immunophenotyping technique whereby sing cell suspensions are stained for either cell surface markers or intracellular proteins by fluorescently-labelled antibodies and analyzed with a flow cytometer, where fluorescently-labelled molecules are excited by the laser to emit light at varying wavelengths, which is then detected by the instrument. There are several key criteria which are required to be kept in mind while designing a flow experiment- 1. Antibody titration (optimal dilution of antibodies should be calculated in order to avoid over- or under- saturated signals for proper detection of surface and intracellular markers), 2. Precision (3 or more replicates of the sample should be used per experiment), 3. Specificity (proper isotype controls should be included in the experiment), 4. Day-to-day variability (experiments should be repeated 3 or more times to ensure consistency and avoid variability due to flow cytometer settings), 5. Antibody interaction (Fluorescence minus one or FMO should be used, which is the comparison of signals from panel minus one antibody vs. the full panel), and 6. Antibody stability (fluorescently-labelled antibodies should be stored at 4C).

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Found 3 matching solutions for this experiment

CD133/2 Antibody, anti-human, APC

Miltenyibiotec

Upstream tips	Protocol tips	Downstream 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		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).

Manufacturer protocol Publication protocol 1 Relevant paper

CD133/2 Antibody, anti-human, PE

Miltenyibiotec

Upstream tips	Protocol tips	Downstream tips
	>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer.

Protocol tips
>20 × 105 disaggregated sphere cells were stained with a stem cell‐specific antibody panel and analysed on a BDTM LSR II flow cytometer. To a volume of 100 μL cell suspension, 5 μL antibody mix was added, mixed and incubated for 20 minutes at 4°C in the dark and washed twice with 250 μL DPBS or FACS buffer.

Manufacturer protocol Publication protocol 1 Relevant paper

CD133 (Prominin-1) Monoclonal Antibody (13A4), APC, eBioscience™

eBioscience

Upstream tips	Protocol tips	Downstream tips
	cells were incubated on ice for 30min with appropriate antibodies or isotype controls washed twice with FACS buffer (PBS containing 0.1% NaN3 and 5% fetal bovine serum), and resuspended in 0.5 ml 1% formalin/PBS.	All analyses were performed on a FACS calibur system (Becton Dickinson Immunocytometry Systems).

Protocol tips
cells were incubated on ice for 30min with appropriate antibodies or isotype controls washed twice with FACS buffer (PBS containing 0.1% NaN3 and 5% fetal bovine serum), and resuspended in 0.5 ml 1% formalin/PBS.
Downstream tips
All analyses were performed on a FACS calibur system (Becton Dickinson Immunocytometry Systems).

Manufacturer protocol Publication protocol 1 Relevant paper

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