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K63 ubiquitin

Manufactured by Merck Group
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The K63 Ubiquitin is a lab equipment product manufactured by Merck Group. It is a protein used in research applications. The core function of the K63 Ubiquitin is to serve as a tool for studying cellular processes involving protein modifications.

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Lab products found in correlation

P mkk4, by Cell Signaling Technology (1 mentions) K48 ubiquitin, by Merck Group (1 mentions) P jnk, by Cell Signaling Technology (1 mentions) β tubulin, by Developmental Studies Hybridoma Bank (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Poly 1 c hmw, by InvivoGen (1 mentions) Ifn β enzyme linked immunosorbent assay elisa kit, by PBL Assay Science (1 mentions) 2 3 cgamp, by InvivoGen (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Anti flag m2 antibody, by Merck Group (1 mentions) Hek293t, by American Type Culture Collection (1 mentions) Vectashield, by Thermo Fisher Scientific (1 mentions) Hsp60, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 568 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Prolongtm glass antifade mountant, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)

4 protocols using k63 ubiquitin

1

Antibody Detection for Ubiquitin-Proteasome System

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The following antibodies were used: USP14 [9 (link)]; β-tubulin (Developmental Studies Hybridoma Bank, Iowa City, IA); Rpt1 and MLK3 (Santa Cruz Biotechnologies, Dallas, TX); Ubiquitin (UAB Hybridoma Facility, Birmingham, AL), K48 Ubiquitin and K63 Ubiquitin (Millipore, Billerica, MA); pMKK4, MKK4, pJNK, and JNK (Cell Signaling Technology, Danvers, Massachusetts).
Vaden J.H., Bhattacharyya B.J., Chen P.C., Watson J.A., Marshall A.G., Phillips S.E., Wilson J.A., King G.D., Miller R.J, & Wilson S.M. (2015). Ubiquitin-specific protease 14 regulates c-Jun N-terminal kinase signaling at the neuromuscular junction. Molecular Neurodegeneration, 10, 3.
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2

Murine Embryonic Fibroblast Cell Lines

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Murine Embryonic Fibroblast (MEF) cell lines were kindly provided by Dr. Genhong Cheng (University of California, Los Angeles). U937 human monocytes and Human Embryonic Kidney (HEK) 293T cells were obtained from American Type Culture Collection (ATCC). Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Corning) (MEF and HEK 293T) or Roswell Park Memorial Institute 1640 (RPMI 1640) (Corning) (U937) supplemented with fetal bovine serum (Gibco) (10%) and penicillin/streptomycin (Gibco) (1%) in 5% CO2 at 37°C. Poly (I:C) (HMW) and 2’3’-cGAMP were purchased from Invivogen. Calf-thymus DNA and oligos for ISD generation were synthesized by Invitrogen. Recombinant murine IFN-β was purchased from MedChemExpress. An IFN-β enzyme linked immunosorbent assay (ELISA) kit were from PBL Assay Science. Anti-Flag M2 antibody was purchased from Sigma-Aldrich. Other primary antibodies used in this study were GFP, HA, Myc, VSV-G, HSV-1 ICP4 (Santa Cruz Biotechnology); TBK1, p-TBK1, IRF3, p-IRF3, STAT1, p-STAT1 (Cell Signaling Technology); TRIM28, β-Tubulin, GAPDH, RSAD2, control IgG (Proteintech); and K63 ubiquitin (Millipore). Secondary anti-rabbit, anti-mouse, or anti-rabbit Trueblot antibodies conjugated to HRP were purchased from Southern Biotech and Rockland Immunochemicals, respectively.
Hua F., Nass T J.r, & Parvatiyar K. (2024). TRIM28 facilitates type I interferon activation by targeting TBK1. Frontiers in Immunology, 15, 1279920.
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3

Immunofluorescence Staining of Mitochondrial Proteins

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Cells were fixed using 4% PFA for 15 min, followed by a 15 min permeabilization step using 0.2% Triton X-100. Samples were blocked using 2% BSA in PBS for 1 h and incubated with primary antibody in 2% BSA overnight. The following day, samples were incubated with a secondary antibody in 2% BSA. Primary antibodies used are COXIV (CST #11967 and #4850), cytochrome c (BD #556432), FK2 (ENZO #BML-PW8810-0100), HSP60 (Santa Cruz #sc-13115), K63-ubiquitin (Merck #05-1308), p65 (CST #8242), TOM20 (CST #42406 and Proteintech #11082-1-AP) and UBCJ2 (ENZO #ENZ-ABS840-0100). Secondary antibodies used are Alexa Fluor 488 goat anti-rabbit IgG (H + L) (Invitrogen #A11034), Alexa Fluor 488 goat anti-mouse IgG (H + L) (Invitrogen #A11029), Alexa Fluor 568 goat anti-rabbit IgG (H + L) (Invitrogen #A11011), Alexa Fluor 568 goat anti-mouse IgG (H + L) (Invitrogen #A11004), Alexa Fluor 647 goat anti-rabbit IgG (H + L) (Invitrogen #A21245) and Alexa Fluor 647 goat anti-mouse IgG (H + L) (Invitrogen #A21236). Coverslips were mounted using Vectashield or ProLongTM Glass Antifade Mountant (Invitrogen #P36980) with or without DAPI.
Vringer E., Heilig R., Riley J.S., Black A., Cloix C., Skalka G., Montes-Gómez A.E., Aguado A., Lilla S., Walczak H., Gyrd-Hansen M., Murphy D.J., Huang D.T., Zanivan S, & Tait S.W. (2024). Mitochondrial outer membrane integrity regulates a ubiquitin-dependent and NF-κB-mediated inflammatory response. The EMBO Journal, 43(6), 904-930.
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4

DNA damage response in human cell lines

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HeLa, U2OS and HEK293Tcells were obtained from the American Type Culture Collection (ATCC; Rockville, MD) were cultured in DMEM (Invitrogen) supplemented with 10% FBS (HyClone) as recommended. H2AX+/+ and H2AX−/− MEFs were a gift from Andre Nussenzweig, and MDC1 proficient and deficient MEFs were a gift from Zhenkun Lou. GM00200 (ATM + / + ) and GM09607 (ATM −/−_) were obtained from Coriel. Cells were grown at 37 °C and in a humidified chamber with 5% CO2.Drugs and Antibodies: G9a inhibitor UNC0638 (Sigma-Aldrich), ATM kinase inhibitor KU-55933 (Astra-Zeneca/Kudos Pharmaceuticals (Cambridge, United Kingdom), 0.5 μM), Neocarzinostatin (NCS, Sigma Aldrich, 200 ng/ml,), and ATR inhibitor VE-821 (Cayman chemicals, 3 μM). Antibodies used were: G9a (Cell Signaling Technology), GLP1 (Bethyl Laboratories), phosphorylated (Ser139) H2AX (EMD Millipore), 53BP1 (Bethyl Laboratories), BRCA1 (Gift from Bing Xa), BARD1 (Santa Cruz Biotechnology), anti-PAR (Travigen), SUMO (Santa Cruz Biotechnology), H3K9me2 (EMD millipore), Flag (Sigma Aldrich), SPOC1 (Sigma Aldrich), K63-ubiquitin (EMD Millipore), GFP (Abcam) and GAPDH (Abcam), For immunofluorescence experiments, the secondary antibodies used were: anti-mouse (115-035-068, 1:20,000) and anti-rabbit (711-035-152, 1:20,000) from Jackson ImmunoResearch Labs.
Ginjala V., Rodriguez-Colon L., Ganguly B., Gangidi P., Gallina P., Al-Hraishawi H., Kulkarni A., Tang J., Gheeya J., Simhadri S., Yao M., Xia B, & Ganesan S. (2017). Protein-lysine methyltransferases G9a and GLP1 promote responses to DNA damage. Scientific Reports, 7, 16613.
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