Phenotype of cultured cells was analyzed using directly labeled monoclonal mouse antibodies: PE-conjugated anti-CD133 and FITC-conjugated anti-CD271 (Miltenyi Biotec, Germany); PE-conjugated anti-CD90 (BD Biosciences, USA); PE-conjugated anti-CD105, PE-conjugated anti-HLA-DR (BD Pharmingen) and anti-CD146 (R&D Systems, USA). All antibodies were used at the concentration of 1 to 5 µl of antibody for 1 × 105 cells in 50 µl of PBS supplemented with 1% FBS, as suggested by the manufacturer, and incubated for 30 min at 4 °C. After incubation cells were carefully washed. IgG isotype matched, PE or FITC labeled immunoglobulins were used as controls. Cells were analyzed by flow cytometry using FACSCalibur (Becton Dickinson, CA). Data were presented using WinMDI 2.7 software.
Anti cd146
Manufactured by R&D Systems
Sourced in United States
Anti-CD146 is a laboratory research tool used to detect and analyze CD146, a cell surface protein. It is commonly used in flow cytometry and immunohistochemistry applications to identify and characterize CD146-expressing cells. The product provides a reliable and specific means of studying CD146 expression in various research contexts.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd90, by R&D Systems (2 mentions)
Anti cd44, by R&D Systems (2 mentions)
Pe conjugated anti cd90, by BD (1 mentions)
Pe conjugated anti cd105, by BD (1 mentions)
Pe conjugated anti hla dr, by BD (1 mentions)
Facscalibur, by BD (1 mentions)
Anti cd45, by R&D Systems (1 mentions)
Anti cd105, by R&D Systems (1 mentions)
Canto 2, by BD (1 mentions)
Anti stro 1, by R&D Systems (1 mentions)
Paraformaldehyde (pfa), by Fujifilm (1 mentions)
Citrate buffer, by Merck Group (1 mentions)
Prolong gold antifade, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal microscope, by Olympus (1 mentions)
Fluoview software, by Olympus (1 mentions)
4 protocols using anti cd146
1
Immunophenotyping of Cultured Cells
2
Phenotypic Characterization of MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MSCs were incubated with anti-CD19, anti-CD146, anti-CD44, anti-CD45, anti-CD90, and anti-CD105 antibodies (R&D, USA) at room temperature for 30 min. After washing twice with PBS, the MSCs were incubated with a FITC-labeled secondary antibody in the dark for 30 min. After washing, the cells were suspended in PBS and analyzed on a flow cytometer.
3
Stem Cell Immunophenotyping by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The percentage of surface markers indicating the stem cell immunophenotype was analyzed by flow cytometry. The PDLSCs, PULPSCs, BMSCs, and CDCs from each group were harvested in PBS, washed once with PBS, fixed in 3.7% paraformaldehyde (Wako, Tokyo, Japan), and then incubated with anti-Stro-1, anti-CD44, anti-CD90, and anti-CD146, antibodies (R&D Systems, Minneapolis, MN, USA) at a dilution of 1:100 (volume/volume) in PBS for 1 hour at room temperature in the dark. Immunostaining with an antibody against the hematopoietic marker anti-CD19 and anti-CD45 was conducted as a negative control. The cells were then incubated with secondary anti-mouse antibodies—allophycocyanin-conjugated antibody for CD44, CD90, CD146, CD19 and CD45 or phycoerythrin-conjugated antibody for STRO-1—at a dilution of 1:100 (volume/volume) in PBS for 30 minutes at 4°C in the dark, and the staining was finally analyzed with a flow-cytometry cell-sorting device (CANTO II, Becton Dickinson, Franklin Lakes, NJ, USA) [30 (link)].
4
Characterizing Clear Cell Renal Carcinoma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Clear cell renal carcinoma tissue analysis was performed as follows. Sections were deparaffinized, hydrated and then incubated in Citrate Buffer (SIGMA) for Antigen Retrieval at 96 °C water bath for 10 min. Tissues were washed in PBS and incubated with Glycine 1 M for 1 h. Sections were incubated with primary antibody anti-EpCAM (Dako Agilent) (1: 50) and anti CD146 (1: 50) (R&D) in PBS containing 3% BSA, 3% FBS and 0,1% Triton X-100 overnight at 4 °C. After washing in PBS, sections were incubated with a mouse alexa 555 and goat alexa 647-conjugated secondary antibody [1: 500 (Thermo Fisher) in PBS containing 3% BSA, 3% FBS and 0,1% Triton X-100] 1 h at 37 °C and then stained for 15 min with DAPI (Invitrogen) diluted in PBS 3% BSA, and subsequently mounted with Prolong-Gold antifade (Invitrogen). Slides were analyzed on a FV1000 Confocal microscope (Olympus, Tokyo, Japan) equipped with 40 × and 60 × oil immersion objectives by the Olympus Fluoview software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!