Lysates from primary melanocytes were purchased from Sciencell Research Laboratories (Carlsbad, CA) and NCI-60 validated human melanoma cell lines obtained from the University of Hawaii Cancer Center included SK-Mel2, SK-Mel28, and MALME-3M. These cell lines were cultured in RPMI media with 10% fetal bovine serum and 1% Antibiotic-Antimycotic (all from GIBCO/Thermo Fisher). Primary antibodies for western blots included rabbit monoclonal anti-SELENOK (Epigemonics, Inc., custom antibody), anti-IP3R1 (Santa Cruz Biotechnology, sc-271197), anti-GAPDH (Santa Cruz Biotechnology, sc-47724), anti-Prom1 (Cell Signaling, 58605). Antibodies from Abcam included anti-TRP2 (ab74073), anti-calcineurin A and B (ab137335, ab154650), and anti-calmodulin (ab 105498, respectively). Western blot secondary antibodies were purchased from Li-Cor Technologies and immunofluorescence secondary Alexafluor595 antibody from Thermo. The transgene encoding full-length SELENOK in pcDNA3.1+ (Invitrogen) has been previously described [11 (link)].
Ab137335
Manufactured by Abcam
Sourced in United States
Ab137335 is a laboratory reagent. It is a primary antibody that can be used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Anti p62, by Cell Signaling Technology (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Ab74073, by Abcam (1 mentions)
Alexafluor 595, by Thermo Fisher Scientific (1 mentions)
Ab154650, by Abcam (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti ip3r1, by Santa Cruz Biotechnology (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
Ab52604, by Abcam (1 mentions)
Glass slide, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 labeled donkey anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
4 protocols using ab137335
1
Evaluation of SELENOK in Melanoma Cells
2
Western Blot Analysis of Cellular Proteins
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Western blot analysis was performed as described previously (Lee et al., 2015 (link)). Cells were lysed using RIPA lysis buffer and whole cell protein was collected. Protein were separated by electrophoresis on a 10–15% sodium dodecyl sulfate polyacrylamide gel and transferred to a nitrocellulose membrane. Immunoreactivity was visualized through sequential incubation with primary antibodies, horseradish peroxidase-conjugated secondary antibodies, and enhanced chemiluminescence reagents. The antibodies used for immunoblotting were anti-calcineurin (ab137335; Abcam), anti-P62 (#5114; Cell Signaling Technology), LC3 (Novus Biologicals, Littleton, CO, USA), PrPC (ab52604, Abcam), and anti-β-actin (A5441; Sigma-Aldrich, St. Louis, MO, USA).
3
Western Blot Analysis of Cell Signaling
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Western blot analysis was measured as described previously [26] . The antibodies used for immunoblotting were anti-calcineurin (ab137335; Abcam), p-nf kb p65(Ser536) (#3033; Cell Signaling Technology, Danvers, MA, USA), p-bcl10, anti-P62 (#5114; Cell Signaling Technology), LC3 (Novus Biologicals, Littleton, CO, USA), and anti-β-actin (A5441; Sigma-Aldrich, St. Louis, MO, USA).
4
Immunocytochemical Analysis of Neuroblastoma
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Immunocytochemical analyses were performed on neuroblastoma cells with p-nfkb (MAB3026; Milipore, Burlington, MA, USA) and anti-calcineurin (ab137335; Abcam). Cells were cultured on glass slides (Nalge Nunc International, Naperville, IL, USA), then washed in sterilized TBST for 10 min, blocked for 15 min with 5% FBS in TBST, and incubated overnight at 4 °C with the primary antibodies diluted with 5% FBS in TBST. Alexa Fluor 488-labeled donkey anti-rabbit IgG antibody, diluted 1:1000 (A21206; Molecular Probes, Eugene, OR, USA), was used to visualize expression using uorescence microscopy.
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