Flow cytometry Anti-bodies Mouse - CD105

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 2 matching solutions for this experiment

Anti-CD105 antibody [MJ7/18] (PE) (ab93567)

Abcam

Upstream tips	Protocol tips	Downstream tips
The first passages of ASCs were trypsinized (1% trypsin-EDTA, Sigma-Aldrich) and then centrifuged at 1,000 ×g for 5 min in the presence of 1% FBS to quench the enzyme. The cell pellets were resuspended in 1% FBS in PBS, filtered through a 70 μm cell strainer (BD Biosciences), and the cells were counted using a One Cell Counter (Wako Chemical).	10 μL anti-CD105 antibody [MJ7/18] (phycoerythrin; ab93567, Abcam) per 1,000,000 cells for 40 min. Antibody conjugated cells were subsequently filtered again through a 35 μm cell strainer (BD Biosciences). Cells were incubated with 5 μL 7-AAD staining solution (BD Biosciences) per 106 cells for 10 min and then sorted according to the expression of the cell surface markers

Upstream tips
The first passages of ASCs were trypsinized (1% trypsin-EDTA, Sigma-Aldrich) and then centrifuged at 1,000 ×g for 5 min in the presence of 1% FBS to quench the enzyme. The cell pellets were resuspended in 1% FBS in PBS, filtered through a 70 μm cell strainer (BD Biosciences), and the cells were counted using a One Cell Counter (Wako Chemical).
Protocol tips
10 μL anti-CD105 antibody [MJ7/18] (phycoerythrin; ab93567, Abcam) per 1,000,000 cells for 40 min. Antibody conjugated cells were subsequently filtered again through a 35 μm cell strainer (BD Biosciences). Cells were incubated with 5 μL 7-AAD staining solution (BD Biosciences) per 106 cells for 10 min and then sorted according to the expression of the cell surface markers

Manufacturer protocol Publication protocol 1 Relevant paper

Purified anti-mouse CD105 Antibody

BioLegend

Upstream tips	Protocol tips	Downstream tips
Each cell type was single cell suspended and, incubated in FACS buffer containing pre-conjugated antibodies (see supplementary Table 2) for 20 min on ice.	. Following incubation, cells were washed twice with FACS buffer.	All the flow cytometry performed using a Cell Lab Quanta SCMPL. The data were analyzed using Kaluza®1.2 software (Beckman Coulter, Denmark) and the expression of each CD markers on the cells was calculated based on the percentage and mean fluorescence intensity (MFI).

Upstream tips
Each cell type was single cell suspended and, incubated in FACS buffer containing pre-conjugated antibodies (see supplementary Table 2) for 20 min on ice.
Protocol tips
. Following incubation, cells were washed twice with FACS buffer.
Downstream tips
All the flow cytometry performed using a Cell Lab Quanta SCMPL. The data were analyzed using Kaluza®1.2 software (Beckman Coulter, Denmark) and the expression of each CD markers on the cells was calculated based on the percentage and mean fluorescence intensity (MFI).

Manufacturer protocol Publication protocol 1 Relevant paper

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