Antibodies for flow cytometry were purchased from Cell Signaling Technologies (pY397FAK #3283, pPaxilin #2541), abcam (TET3 #ab135033, 5hmU #ab19735), Novus Biologicals (laminin #NB300–144), Active Motif (5mC #39649) or BD Biosciences (pSTAT3 #557815). Secondary antibodies used were coupled to either Alexa Fluor 647 or Alexa Fluor 488 (Invitrogen) diluted up to 1:4,000 in PBS/1% BSA staining buffer. For blocking FAK activity, GSC cell lines were treated with 10 μM FAK inhibitor (Santa Cruz #sc-203950). Intracellular staining was performed after fixation of single cell suspensions with 2% paraformaldehyde and permeabilization with ice-cold 100% methanol, blocked with PBS/1% BSA for 1h at 4°C, and stained with an antibody diluted at 1:50 in PBS/1% BSA for 30 – 45 min at room temperature. Detection of DNA modification was performed upon RNA digestion using RNase A at 40 μg/ml for 10 min at 37°C prior to blocking procedure. Stained single cell suspensions were analyzed using an AccuriC6 cytometer (BD Biosciences), followed by FlowJo 7.6.1 analysis.
Sc 203950
Manufactured by Santa Cruz Biotechnology
The Sc-203950 is a laboratory instrument designed for general use in biotechnology and life science research applications. It is a standard piece of equipment commonly found in research laboratories. The core function of this product is to facilitate the analysis and processing of biological samples. Detailed technical specifications are not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Laminin nb300 144, by Active Motif (2 mentions)
Pstat3 557815, by BD (2 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Accuri c6 cytometer, by BD (2 mentions)
Ab23680, by Abcam (1 mentions)
Ab52968, by Abcam (1 mentions)
Latex bead, by Merck Group (1 mentions)
Cathepsin d activity assay kit, by Abcam (1 mentions)
A5316, by Merck Group (1 mentions)
Arg gly asp rgd peptide, by Merck Group (1 mentions)
Sc 10804, by Santa Cruz Biotechnology (1 mentions)
Sc 28259, by Santa Cruz Biotechnology (1 mentions)
3 protocols using sc 203950
1
Antibody-based Flow Cytometry Analysis
2
Antibody-mediated phagocytosis in ARPE-19 cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The following antibodies were used in this study: ZO-1 (1:100; sc-10804, Santa Cruz Biotechnology, Santa Cruz, CA, USA), prohibitin (1:100; sc-28259, Santa Cruz), transcriptional factor EB (TFEB) (1:100; sc-11004, Santa Cruz), CD36 (2 μg/ml; ab23680, Abcam, Cambridge, MA, US), MerTK (3.44 μg/ml; ab52968, Abcam), PGC-1α antibody (1μg/ml; ST1202, Calbiochem, La Jolla, CA, USA), and β-actin (1:2000; A5316, Sigma-Aldrich, St. Louis, MO, USA). ARPE-19 cells were pretreated with anti-CD36 and anti-MerTK antibodies for 1 h before a 3-h POS treatment. Latex beads (0.6 μm, 10 beads/cell, LB6) and Arg–Gly–Asp (RGD) peptide (0.5 mM; A8052) were from Sigma–Aldrich. Further, ARPE-19 cells were pretreated with RGD peptide for 30 min before a 3-h POS treatment. The cells were also pretreated with focal adhesion kinase (FAK) inhibitor 14 (500 μM) (sc-203950, Santa Cruz) for 30 min. Cathepsin D activity assay kit (ab65302) was purchased from Abcam.
3
Antibody-based Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies for flow cytometry were purchased from Cell Signaling Technologies (pY397FAK #3283, pPaxilin #2541), abcam (TET3 #ab135033, 5hmU #ab19735), Novus Biologicals (laminin #NB300–144), Active Motif (5mC #39649) or BD Biosciences (pSTAT3 #557815). Secondary antibodies used were coupled to either Alexa Fluor 647 or Alexa Fluor 488 (Invitrogen) diluted up to 1:4,000 in PBS/1% BSA staining buffer. For blocking FAK activity, GSC cell lines were treated with 10 μM FAK inhibitor (Santa Cruz #sc-203950). Intracellular staining was performed after fixation of single cell suspensions with 2% paraformaldehyde and permeabilization with ice-cold 100% methanol, blocked with PBS/1% BSA for 1h at 4°C, and stained with an antibody diluted at 1:50 in PBS/1% BSA for 30 – 45 min at room temperature. Detection of DNA modification was performed upon RNA digestion using RNase A at 40 μg/ml for 10 min at 37°C prior to blocking procedure. Stained single cell suspensions were analyzed using an AccuriC6 cytometer (BD Biosciences), followed by FlowJo 7.6.1 analysis.
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