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Sc 81514

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Sc-81514 is a laboratory product from Santa Cruz Biotechnology. It is a chemical reagent used in biological research applications.

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Lab products found in correlation

Nocodazole, by Selleck Chemicals (1 mentions) Ab81292, by Abcam (1 mentions) Ab201022, by Abcam (1 mentions) Ku55933, by Selleck Chemicals (1 mentions) Ab183479, by Abcam (1 mentions) Ab70385, by Abcam (1 mentions) Lambda protein phosphatase λppase, by Merck Group (1 mentions) Sc 9996, by Santa Cruz Biotechnology (1 mentions) Sc 5267, by Santa Cruz Biotechnology (1 mentions) F3165, by Merck Group (1 mentions) Streptavidin agarose beads, by GE Healthcare (1 mentions) Pan phospho serine antibody, by Santa Cruz Biotechnology (1 mentions) Pka kinase activity assay kit, by Abcam (1 mentions)

2 protocols using sc 81514

1

Immunoblotting Analysis of Mitotic Regulators

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Nocodazole and Ku55933 were ordered from Selleckchem (Texas, USA). Lambda Protein phosphatase (λPPase) was bought from Sigma-Aldrich Corporation. The anti-ATM (1:1,000, ab201022, Abcam), anti-phospho-ATM (1:1,000, ab81292, Abcam), anti-Mad2 (1:1,000, ab70385, Abcam), anti-Mad1 (1:1,000, ab201022, Abcam), and anti-Cdc20 antibodies (1:1,000, ab183479, Abcam) were bought from Abcam (Cambridge, MA). The anti-HA (1:1,000, 3724T, Cell Signaling Technology), anti-Mad2 (1:1,000, 4636S, Cell Signaling Technology), and the HRP-conjugated secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA). The anti-Flag antibody (1:1,000, F3165, Sigma) was purchased from Sigma. The anti-Mad2 antibody (1:1,000, YT2618, Immunoway) was bought from Immunoway. The anti-GFP (1:1,000, sc-9996, Santa Cruz), anti-Mad2 (1:1,000, sc-47747, Santa Cruz), anti-p-Thr (1:1,000, sc-5267, Santa Cruz), and anti-p-Ser antibodies (1:1,000, sc-81514, Santa Cruz) were bought from Santa Cruz Biotechnology. Goat anti-mouse lgG-H HRP (1:1,000, M21004L, Abmart) and goat anti-mouse lgG-L HRP (1:1,000, M21005S, Abmart) were purchased from Abmart.
Wang Y., Yu T., Han Y., He Y., Song Y., Guo L., An L., Yang C, & Wang F. (2022). Phosphorylation of MAD2 at Ser195 Promotes Spindle Checkpoint Defects and Sensitizes Cancer Cells to Radiotherapy in ATM Deficient Cells. Frontiers in Cell and Developmental Biology, 10, 817831.
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2

TFAM Phosphorylation by PKA Kinase Assay

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Hydroxylated TFAM (1 μg) biotinylated peptide was conjugated with
30 μL Streptavidin agarose beads (GE Healthcare Life Sciences) in 1 ml
PBS at room temperature with rotation for 1 hour. The beads pellet was washed
three times with PBS and resuspended in 500 μL PBS supplemented with 1
μg purified GST-pVHL protein for 2 hours at room temperature.
Subsequently, TFAM phosphorylation by PKA was assayed using PKA Kinase Activity
Assay Kit (ab139435, Abcam) according to manufacturer's instructions.
Samples were centrifuged at 8000 x g for 30 sec and washed three times with PBS
and re-suspended with 50 μL kinase assay dilution buffer supplemented
with 1 μg/μL ATP and 50 ng purified active PKA. The
phosphorylation assay was processed for 40 min at room temperature with rotation
and for 40 min at 30 °C. Samples were centrifuged at 8000 x g for 30 sec
and washed three times with PBS and eluted with 30 μL Laemmli buffer,
boiled for 5 min and centrifuged at 8000 x g for 30 sec. Eluted supernatant was
analyzed by immunoblotting using pan-phospho-serine antibody (Santa Cruz:
sc-81514).
Li S., Li W., Yuan J., Bullova P., Wu J., Zhang X., Liu Y., Plescher M., Rodriguez J., Bedoya-Reina O.C., Jannig P.R., Valente-Silva P., Yu M., Henriksson M.A., Zubarev R.A., Smed-Sörensen A., Suzuki C.K., Ruas J.L., Holmberg J., Larsson C., Christofer Juhlin C., von Kriegsheim A., Cao Y, & Schlisio S. (2022). Impaired oxygen-sensitive regulation of mitochondrial biogenesis within the von Hippel–Lindau syndrome. Nature Metabolism, 4(6), 739-758.
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