Immunohistochemical staining was performed to detect the expression of proteins in tissues and cells. Anti‐PSMA antibody (1:250; ab133579), anti‐CD56 antibody (1:2000; ab220360), anti‐solute carrier family 3‐member 2 (SLC3A2) antibody (1:2500; ab244356) and anti‐solute carrier family 7‐member 11 (SLC7A11) antibody (1:500; ab37185) were purchased from Abcam. The PSMA, CD56, SLC3A2, and SLC7A11 protein expression levels were assessed by IHC according to the recommended protocol [19 (link)]. Briefly, the protocol included tissue or cell fixation, serum blocking, primary antibody incubation (4°C, 12 hours), marked second antibody incubation (25°C, 30 minutes), staining, judgment of the results and imaging. The DNA dye 4',6‐diamidino‐2‐phenylindole (DAPI) was obtained from Molecular Probes (San Francisco, CA, USA). The secondary antibody used was Cy3‐conjugated donkey anti‐rabbit IgG (Jackson ImmunoResearch Laboratory, West Grove, PA, USA). All stained cells were examined and photographed with a Leica SP5 confocal fluorescence microscope (Wetzlar, Germany).
Ab244356
Manufactured by Abcam
Ab244356 is a lab equipment product available from Abcam. It serves a core function as a laboratory tool, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Sp5 confocal fluorescence microscope, by Leica (1 mentions)
Cy3 conjugated donkey anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Ab133579, by Abcam (1 mentions)
Ab220360, by Abcam (1 mentions)
Ab37185, by Abcam (1 mentions)
Dna dye 4 6 diamidino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions)
Proteinase inhibitor, by Beyotime (1 mentions)
Ab40794, by Abcam (1 mentions)
2 protocols using ab244356
1
Immunohistochemical Analysis of Protein Expression
2
Western Blot Analysis of Protein Markers
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Proteins were extracted with radioimmunoprecipitation buffer containing proteinase inhibitor (Beyotime), and then separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) (30 μg per sample) before nitrocellulose membrane transfer (Millipore Biotech, MA, USA). The membranes were blocked with 5% skim milk and probed with primary and horseradish peroxidase (HRP)-conjugated rabbit secondary antibodies (Beyotime). To detect the signal, an enhanced chemiluminescence system (ECL, Millipore Biotech) was used. Band density was quantified using ImageJ software (http://rsb.info.nih.gov/ij/ , MD, USA) and normalized to GAPDH. The primary antibodies were as follows: EPAS1 (Abcam, ab222396, 1:2000), SLC3A2 (Abcam, ab244356, 1:1000), p-AKT (CST, #4060, 1:1000), AKT (CST, #4685, 1:1000), p Y576-FAK (Abcam, ab76120, 1:1000), FAK (Abcam, ab40794, 1:500), Cas (Abcam, ab92514, 1:1000), pY20-Cas (BioSource International, AHO0681, 1:1000), cleaved caspase-3 (CST, #9661, 1:1000), and GAPDH (CST, #5174, 1:1000).
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