Antibody incubation was performed on ice for 15 min in PBS with 2% fetal bovine serum. The primary antibodies employed were anti-CD24-PeCy7 (eBiosciences, 25-0291-82, 1:1000), anti-CD24-BV711 (BD Biosciences, 563450, 1:1000), anti-CD29-PeCy7 (eBiosciences, 25-0242-82, 1:1000), anti-CD29-FITC (BD Biosciences, 561796, 1:1000), anti-CD45-APC (eBiosciences, 17-0451-82, 1:1000), anti-CD31-APC (eBiosciences, 17-0311-82, 1:1000), anti-TER119-APC (eBiosciences, 17-5921-82, 1:1000) and anti-Dyeflour450 (eBiosciences, 65-0863-14, 1:1000). Flow cytometry analysis was performed using Aria SROP (BD). The data were analyzed using FlowJo software. For sorting cells, an Aria III (BD) instrument was used.
Anti cd29 fitc
Manufactured by BD
Sourced in United States
Anti-CD29-FITC is a fluorescently-labeled antibody that binds to the CD29 antigen. CD29 is a cell surface protein expressed on various cell types. The FITC label allows for detection and analysis of CD29-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd44 fitc, by BD (2 mentions)
Anti cd31 apc, by Thermo Fisher Scientific (1 mentions)
Anti cd24 pecy7, by Thermo Fisher Scientific (1 mentions)
Anti cd45 apc, by Thermo Fisher Scientific (1 mentions)
Anti ter119 apc, by Thermo Fisher Scientific (1 mentions)
Aria 3, by BD (1 mentions)
Anti cd90 pe, by BD (1 mentions)
Lsrii four laser flow cytometer, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Anti cd45 percp cy5, by BioLegend (1 mentions)
Compbead, by BD (1 mentions)
Facsaria 3, by BD (1 mentions)
Alizarin red s staining, by Solarbio (1 mentions)
Anti cd90 fitc, by BD (1 mentions)
Anti cd31 pe, by BD (1 mentions)
Anti cd105 pe, by BD (1 mentions)
Alcian blue, by Merck Group (1 mentions)
Anti cd45 pe, by BD (1 mentions)
Anti cd73 pe, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
4 protocols using anti cd29 fitc
1
Multiparameter Analysis of Cell Surface Markers
2
Characterization of Mesenchymal Stem Cells
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Flow cytometry was performed by using an LSRII four-laser flow cytometer (BD Biosciences, Oxford, UK) with appropriate isotype controls. Passaged MSCs (p3 from HN, SF, and ICBM) (n = 3 for each group) were stained with combinations of the following antibodies at the dilution recommended by the manufacturers: anti-CD14-allophycocyanin-cyanine (APC-H7), anti-CD19-fluorescein isothiocyanate (FITC), anti-CD34-peridinin chlorophyll protein (PerCP), anti-HLADR-phycoerythrin-cyanine (PE-Cy7), anti-CD73-phycoerythrin (PE), anti-CD90-PE, anti-CD81-FITC, anti-CD44-FITC, and anti-CD29-FITC (all from BD Biosciences). Cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich) as a live/dead discriminator immediately prior to acquisition
[15 (link)]. At least 10,000 live cell events were collected for each antibody combination.
[15 (link)]. At least 10,000 live cell events were collected for each antibody combination.
3
Characterization of Murine Bone Marrow Cells
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Flow cytometry antibodies anti-CD29-FITC, and CompBeads were purchased from BD biosciences (New Jersey, USA). Anti-CD45-PerCPcy 5.5 was from Biolegend (USA) and anti-CD44-APC was from eBioscence (SanDiego, CA, USA). Bone marrow cells from tibia and femur of 10-12-week-old saline or CPD-treated (7 consective days, dosages) male mice were flushed out with PBS plus 2.5% Fetal bove serum (FBS), filtered, centrifuged, removed the red blood cell, and collected. The final concentrations were 1×107/ml before performing flow cytometry assays. The negative expression of CD45-PerCPcy5.5, and positive expression of CD29-FITC, and CD44-APC were obtained and analyzed with BD FACS Aria III (New Jersey, USA), and one representative assay was shown.
4
Characterization of ADSC phenotype and multilineage differentiation
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To identify ADSC surface markers, ADSCs at passage 3 were collected in tubes at 4 × 105/tube. Next, anti-CD31-PE, anti-CD45-PE, anti-CD44-FITC, anti-CD29-FITC, anti-CD73-PE, anti-CD90-FITC, and anti-CD105-PE (all from BD Biosciences, San Jose, CA, USA) antibodies were incubated. Then, ADSCs in solution were identified by a FACS Calibur flow cytometer (Becton-Dickinson, Franklin Lakes, NJ, USA).
Trilineage differentiation was performed to show the differentiation potency of ADSCs. In brief, ADSCs were seeded into 6-well plates (5.0 × 105 cells/well) with normal medium until cell confluence reached approximately 70%. The nonadherent cells were removed by replacing the medium, and the attached cells were cultured until confluence. The cells were then grown for 21 days in the adipogenic, osteogenic, and chondrogenic medium (Cyagen, Guangzhou, China). Alizarin red S staining (ARS, Solarbio, Beijing, China) was performed to assess osteogenic differentiation. Adipogenic differentiation was visualized using Oil Red O (Sigma-Aldrich, St. Louis, MO, USA) staining. Alcian Blue (Sigma-Aldrich, St. Louis, MO, USA) staining was used to assess chondrogenic differentiation.
Trilineage differentiation was performed to show the differentiation potency of ADSCs. In brief, ADSCs were seeded into 6-well plates (5.0 × 105 cells/well) with normal medium until cell confluence reached approximately 70%. The nonadherent cells were removed by replacing the medium, and the attached cells were cultured until confluence. The cells were then grown for 21 days in the adipogenic, osteogenic, and chondrogenic medium (Cyagen, Guangzhou, China). Alizarin red S staining (ARS, Solarbio, Beijing, China) was performed to assess osteogenic differentiation. Adipogenic differentiation was visualized using Oil Red O (Sigma-Aldrich, St. Louis, MO, USA) staining. Alcian Blue (Sigma-Aldrich, St. Louis, MO, USA) staining was used to assess chondrogenic differentiation.
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