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K63 ubiquitin

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K63-ubiquitin is a specific type of ubiquitin protein that is conjugated to target proteins, primarily through a lysine 63 linkage. Ubiquitin is a small regulatory protein that plays a crucial role in various cellular processes by attaching to other proteins, thereby modulating their function, localization, or stability.

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Lab products found in correlation

K48 ubiquitin, by Cell Signaling Technology (3 mentions) Bca kit, by Thermo Fisher Scientific (3 mentions) Biotin, by Cell Signaling Technology (2 mentions) Nedd8, by Cell Signaling Technology (2 mentions) Hnrnp e1, by Abnova (2 mentions) Hsp90, by Santa Cruz Biotechnology (2 mentions) Flag tag (flag), by Cell Signaling Technology (2 mentions) E cadherin, by Cell Signaling Technology (2 mentions) Vimentin, by Cell Signaling Technology (2 mentions) Chemidoc system, by Bio-Rad (2 mentions) N cadherin, by BD (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Ubiquitin, by Cell Signaling Technology (1 mentions) Z vad fmk, by BD (1 mentions) Lamp2, by Santa Cruz Biotechnology (1 mentions) P tbk1, by Cell Signaling Technology (1 mentions) Mg132, by Merck Group (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions)

6 protocols using k63 ubiquitin

1

Immunoblot Analysis of Protein Interactions

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Immunoblot analysis was performed as described previously 47 (link). Briefly, cells were lysed in RIPA buffer and denatured at 95°C for 5 min in Laemmli buffer. Samples were resolved on SDS–PAGE gels and transferred to PVDF membrane. Blots were probed with antibodies specific to ARIH1 (VPA00397; BioRad), hnRNP E1 (M01; Abnova), E-cadherin (3195; Cell Signaling), N-cadherin (BD Transductions), Vimentin (5741; Cell Signaling), Biotin (5597; Cell Signaling), Flag (14793; Cell Signaling), K48 Ubiquitin (8081; Cell Signaling), K63 Ubiquitin (5621; Cell Signaling), Nedd8 (2745; Cell Signaling), Hsp90 (sc13119: Santa Cruz Biotechnology), CD44 (GTX102111; GeneTex), V5 (R96025; ThermoFisher), UPF1 (VMA00627; BioRad), Filamin-A (VMA00322; BioRad), Cortactin (VMA00430; BioRad), CAND1 (VMA00610; BioRad), and FASN (VMA00266; BioRad). Chemiluminescence was detected by CCD camera (BioRad ChemiDoc system).
Howley B.V., Mohanty B., Dalton A., Grelet S., Karam J., Dincman T, & Howe P.H. (2022). The ubiquitin E3 ligase ARIH1 regulates hnRNP E1 protein stability, EMT and breast cancer progression. Oncogene, 41(12), 1679-1690.
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2

Apoptosis and Autophagy Regulation Assay

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The following chemicals were used for the treatment of cells: general caspase inhibitor Z-VAD-FMK (BD Biosciences, #550377), proteasome inhibitor MG132 (Sigma-Aldrich, C2211), lysosome inhibitor bafilomycin A1 (Sigma-Aldrich, B1793), and a synthetic analog of double-stranded RNA, poly-inosinic-cytidylic acid-high molecular weight (poly I:C-HMW Invivogen, #tlrl-pic). The following primary antibodies were used for western blot analysis and/or immunofluorescence staining: CALCOCO2/NDP52 (Santa Cruz Biotechnology, sc-376540), SQSTM1/p62 (PROGEN Biotechnik GmbH, GSQSTM1-C), cleaved caspase-3 (Cell Signaling Technology, #9661), LC3 (MBL International, PM036), VP1 (Dako, M706401-1), NBR1 (Santa Cruz Biotechnology, sc-130380), HA (Roche, 11867423001), Flag (Sigma, F1804), ubiquitin (Cell Signaling Technology, #3933), GFP (Life Technologies, A-6455), K63-ubiquitin (Cell Signaling Technology, #12930), K48-ubiquitin (Cell Signaling Technology, #12805), MAVS (Cell Signaling Technology, #24930), p-TBK1 (Cell Signaling Technology, #5483), TBK1 (Cell Signaling Technology, #3504), LAMP2 (Santa Cruz Biotechnology, sc-8100), ACTB (Sigma-Aldrich, A5316), and GAPDH (Cell Signaling Technology, 14C10).
Mohamud Y., Qu J., Xue Y.C., Liu H., Deng H, & Luo H. (2018). CALCOCO2/NDP52 and SQSTM1/p62 differentially regulate coxsackievirus B3 propagation. Cell Death and Differentiation, 26(6), 1062-1076.
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3

Immunoblotting of NF-κB Signaling

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The cells were treated as indicated and were prepared in the RIPA lysis buffer (Cell Signaling Technology, Danvers, MA, USA, #9806) and quantified using a BCA kit (Thermo, Waltham, MA, USA, #23227). The following primary antibodies were used: PI16 (Proteintech, Rosemont, IL, USA, #12267-1-AP), NEMO (#ab178872), HOIL-1 (#ab108479) (Abcam, Cambridge, MA, USA), p-IKKα/β (#2697), IKKα (#11930), IKKβ (#8943), IκBα (#4814), p-IκBα (#2859), p65 (#8242), p-p65 (#4887), GAPDH (#5174), p84 (#131268), K63-ubiquitin (#12930), ANXA1 (#32934), HOIP (#99633), Sharpin (#12541) (Cell Signaling) and M1-ubiquitin (Millipore, #MABS451). The secondary antibody (Cell Signaling, #7074, #7076) was probed at the indicated time points. The immunoreactive strips were detected using the ECL Ultra Western HRP Substrate (Millipore, #WBULS0100) detection kit.
Kuang X., Zhang Z., Li D., Bao W., Pan J., Zhou P., Chen H., Gao Z., Xie X., Yang C., Zhu G., Zhou Z., Tang R., Feng Z., Zhou L., Feng X., Wang L., Yang J, & Jiang L. (2023). Peptidase inhibitor (PI16) impairs bladder cancer metastasis by inhibiting NF-κB activation via disrupting multiple-site ubiquitination of NEMO. Cellular & Molecular Biology Letters, 28, 62.
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4

Immunoblot Analysis of Cellular Proteins

Cited 1 time
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Immunoblot analysis was performed as described previously [47 (link)]. Briefly, cells were lysed in RIPA buffer and denatured at 95 °C for 5 min in Laemmli buffer. Samples were resolved on SDS–PAGE gels and transferred to PVDF membrane. Blots were probed with antibodies specific to ARIH1 (VPA00397; BioRad), hnRNP E1 (M01; Abnova), E-cadherin (3195; Cell Signaling), N-cadherin (BD Transductions), Vimentin (5741; Cell Signaling), Biotin (5597; Cell Signaling), Flag (14793; Cell Signaling), K48 Ubiquitin (8081; Cell Signaling), K63 Ubiquitin (5621; Cell Signaling), Nedd8 (2745; Cell Signaling), Hsp90 (sc13119: Santa Cruz Biotechnology), CD44 (GTX102111; GeneTex), V5 (R96025; ThermoFisher), UPF1 (VMA00627; BioRad), Filamin A (VMA00322; BioRad), Cortactin (VMA00430; BioRad), CAND1 (VMA00610; BioRad), and FASN (VMA00266; BioRad). Chemiluminescence was detected by CCD camera (BioRad ChemiDoc system).
Howley B.V., Mohanty B., Dalton A., Grelet S., Karam J., Dincman T, & Howe P.H. (2022). The ubiquitin E3 ligase ARIH1 regulates hnRNP E1 protein stability, EMT and breast cancer progression. Oncogene, 41(12), 1679-1690.
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5

TRIM26 Ubiquitination and GPX4 Regulation in Oxidative Stress

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Antibodies and reagents were acquired from designated suppliers:
TRIM26 (ab188017, Abcam), TRIM26 (27013-1-AP, Proteintech), TRIM26 (sc-393832, Santa Cruz Biotechnology), GPX4 (ab125066, Abcam), GPX4 (sc-166570, Santa Cruz Biotechnology), PLK1 (ab189139, Abcam), p-PLK1 (ab155095, Abcam), β-actin (ab8226, Abcam), K48-ubiquitin (ab140601, Abcam), K63-ubiquitin (12930, Cell Signaling Technology), anti-Flag (66008, Proteintech), anti-Myc (16286, Proteintech), p-S/T (61 G, abmart), p-S/T (612549, BD Biosciences), Anti-His (66005, Proteintech), anti-Flag (14793, Cell Signaling Technology), anti-Myc (2276, Cell Signaling Technology), Anti-His (12689, Cell Signaling Technology), anti-HA (3724, Cell Signaling Technology),
MDA (ab27642, Abcam), 4-HNE (ab48506, Abcam), MG132 (S1748, Beyotime), cycloheximide (HY12320, MedChemExpress), Chloroquine (HY-17589A, MedChemExpress), Onvansertib (HY-15828, MedChemExpress), MLN0905 (HY-15155, MedChemExpress), Recombinant Human PLK1 (ab271716, Abcam).
Wang Z., Xia Y., Wang Y., Zhu R., Li H., Liu Y, & Shen N. (2023). The E3 ligase TRIM26 suppresses ferroptosis through catalyzing K63-linked ubiquitination of GPX4 in glioma. Cell Death & Disease, 14(10), 695.
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6

Molecular Mechanisms of Cell Death

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Cefotaxime Sodium Salt, Cefpirome Sulfate, Cephalexin, SP600125, NAC, salicylate and H2O2 were purchased from Wako. Propyl gallate, Ferrostatin-1 and U0126 were purchased from Sigma. z-VAD-fmk was purchased from Peptide Institute. SB203580, antimycin, apocynin, Necrostatin-1, DPQ and Mito-TEMPO were purchased from Santa Cruz. The antibodies used were against phospho-p38, p38, phospho-JNK, JNK, K48- and K63-ubiquitin, AIF and Lamin A/C (Cell Signaling), p62 (MBL), and β-actin (Santa Cruz). cDNAs encoding human p62 was obtained by performing PCR, and was inserted into pcDNA3 with Flag tag plasmid. Flag-p62 C105A/C113A mutant was generated by mutating 105Cys and 113Cys to Ala in p62. Plasmid transfection was performed using Polyethylenimine “Max” (PEI-MAX, Polysciences), according to the manufacturer’s instructions.
Noguchi T., Suzuki M., Mutoh N., Hirata Y., Tsuchida M., Miyagawa S., Hwang G.W., Aoki J, & Matsuzawa A. (2018). Nuclear-accumulated SQSTM1/p62-based ALIS act as microdomains sensing cellular stresses and triggering oxidative stress-induced parthanatos. Cell Death & Disease, 9(12), 1193.
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