The following antibodies were used for Imm-munofluorescence: PD-L1 (E1LRN, Cell signaling 1:200) and CD8 (CD8/144B, ThermoFisher, 1:200). Primary antibodies were used at the concentration indicated by the manufacturers. Anti-Mouse and anti-Rabbit Alexa488- or Alexa594-conjugated (Life Technologies) were used as secondary antibodies. Alexa594/Alexa488-conjugated antibodies were used at 1:100 dilution. Pictures were acquired using a fluorescence microscope (FL10i, Olympus). Image quantification was performed by ImageJ and analyzed by R-software.
Fl10i
Manufactured by Olympus
The FL10i is a compact and versatile fluorescence microscope designed for a wide range of laboratory applications. It features a powerful LED illumination system and a high-sensitivity camera for capturing detailed fluorescence images. The FL10i is a reliable and user-friendly instrument suitable for various research and analysis tasks.
Automatically generated - may contain errors
3 protocols using fl10i
1
Immunofluorescence Staining of PD-L1 and CD8
2
Immunostaining Analysis of Cellular Signaling
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The following antibodies were used in immunostaining analyses: phospho-JNK (Phospho-SAPK/JNK (Thr183/Tyr185), Cell signaling, 1:200), p-MAPK14 (p-p38 Antikörper (Thr 180/Tyr 182), Santa Cruz 1:200), PD-L1 (E1LRN, Cell signaling 1:200) and phospho-Akt (Phospho-Akt (Thr 308), Cell signaling, 1:200). Primary antibodies were used at the concentration indicated by the manufacturers. Anti-Mouse and anti-Rabbit Alexa488- or Alexa594-conjugated (Life Technologies) were used as secondary antibodies. Alexa594-conjugated antibodies were used at 1:200 dilution and Alexa488-conjugated antibodies were used at 1:100 dilution. Pictures were acquired using a fluorescent microscope (FL10i, Olympus). Image quantification was performed by ImageJ and analyzed by R-software.
3
Antibodies and Immunostaining Protocol
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The following antibodies were used in immunoblotting analyses: phospho-c-Jun (Ser73) (Cell Signaling dilution: 1:500), c-Jun (Cell Signaling dilution: 1:1000), DNMT1 (Abcam dilution: 1:1000), JNK (Santa Cruz, dilution: 1:5000), phospho-JNK (Santa Cruz, dilution: 1:5000), and α-tubulin (mouse monoclonal, Abcam). Primary antibodies were used at the concentration indicated by the manufacturers. Anti-Mouse and anti-Rabbit HRP-conjugated (Santa Cruz, dilution: 1:5000) were used as secondary antibodies. Immunostaining was performed using antibodies against DNMT1 (Abcam dilution: 1:200), CD44 (Abcam dilution: 1:200), CHI3L1 (Quidel dilution: 1:150) and MMP9 (Cell signaling dilution: 1:200). Primary antibodies were used at the concentration indicated by the manufacturers. Anti-Mouse, anti-Rabbit and anti-Goat Alexa594- or Alexa647-conjugated (Life Technologies) were used as secondary antibodies. Alexa594-conjugated antibodies were used at 1:200 dilution and Alexa647-conjugated antibodies were used at 1:100 dilution. Pictures were acquired using a fluorescent microscope (FL10i, Olympus).
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