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Ube2m

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UBE2M is a protein-coding gene that produces an enzyme involved in the ubiquitin-proteasome system. The enzyme catalyzes the covalent attachment of ubiquitin to target proteins, marking them for degradation by the proteasome.

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Lab products found in correlation

Ube2f, by Abcam (2 mentions) Nedd8, by Abcam (2 mentions) Cullin1, by Abcam (2 mentions) Pdnapkcs s2056, by Abcam (1 mentions) Dna pkcs, by Thermo Fisher Scientific (1 mentions) γh2ax, by Merck Group (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Cullin2, by Abcam (1 mentions) Cullin5, by Abcam (1 mentions) C parp, by Cell Signaling Technology (1 mentions) Cullin4b, by Proteintech (1 mentions) Cullin4a, by Cell Signaling Technology (1 mentions) Cullin3, by Cell Signaling Technology (1 mentions) Protease inhibitor, by Beyotime (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) β actin, by Merck Group (1 mentions) Anti myc beads, by Thermo Fisher Scientific (1 mentions) Coro1a, by Abcam (1 mentions) Tsg101, by Abcam (1 mentions)

4 protocols using ube2m

1

DNA-PKcs Regulation by NEDD8 Pathway

Cited 1 time
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All antibodies were purchased from the following sources: NEDD8 (Abcam ab81264), DNA-PKcs (Thermo Fisher Scientific MA5-13238; Abcam ab32566; Proteintech 19983-1-AP), p-DNA-PKcs S2056(Abcam ab18192), Ku70 (Santa cruz sc-17789), Ku80(Santa cruz sc-9034), UBA3(Santa cruz sc-377352), UBE2M(Abcam ab109507), UBE2F(Abcam ab185234), Flag(Sigma F3156), γH2Ax(Millipore 05-636), HUWE1(Bethyl A300-486A-T). SFB-NEDD8, SFB-NEDD8ΔG, SFB-ub, HA-NEDD8, HA-NEDD8 K48R/K60R/NoK constructs were described previously (SFB-tag: S-protein tag, Flag epitope tag and streptavidin-binding peptide tag)13 (link). His-myc-tagged-NEDD8/NEDD8ΔG and NEDP1/NEDP1 C163S constructs were generated by PCR and cloned into pcDNA3.1-Myc-HisB. Flag-tagged-DNA-PKcs-F, M and FATC fragment were cloned into pcDNA3.1. Flag-tagged HUWE1 as previously described22 (link) was gifted from professor Genze Shao (Peking University School of Basic Medical Sciences). His-Myc-tagged-DNA-PKcs-M fragment K to R mutants (1–8) were generated by PCR mutagenesis and then cloned into pcDNA3.1-Myc-HisB. The siRNAs were synthesized by GenePharma (Shanghai China) and used at 10 μm final concentration. siRNA as follows: UBA3(AGAGAGAGAUUAUGAGCAA), UBE2M-1 (CAGAGGUCCUGCAGAACAA), UBE2M-2 (GAUGAGGGCUUCUACAAGA), UBE2F-1 (GGAAUAAAGUGGAUGACUA), UBE2F-2 (CAACAUAAAUACAGCAAGA), HUWE1-1 (CAUUGGAAAGUGCGAGUUA), HUWE1-2 (CUGUGAGAGUGAUCGGGAA).
Guo Z., Wang S., Xie Y., Han Y., Hu S., Guan H., Xie D., Bai C., Liu X., Gu Y., Zhou P.K, & Ma T. (2020). HUWE1-dependent DNA-PKcs neddylation modulates its autophosphorylation in DNA damage response. Cell Death & Disease, 11(5), 400.
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2

Cullin-RING Ubiquitin Ligase Toolkit

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Antibodies specific to C-PARP (Cell Signaling Technology, 5625), Cullin1 (Abcam, 75817), Cullin2 (Abcam, 166917), Cullin3 (Cell Signaling Technology, 2759), Cullin4A (Cell Signaling Technology, 2699), Cullin4B (Proteintech, 12916-1-AP), Cullin5 (Abcam, 184177), NAE (Cell Signaling Technology, 14321), NOXA (Cell Signaling Technology, 14766), P27 (Cell Signaling Technology, 3686), RBX1 (Abcam, 133565), UBA3 (Abcam, 124726), UBE2F (Abcam, 12932), and UBE2M (Abcam, 109507) were purchased commercially.
Zhou L., Zhu J., Chen W., Jiang Y., Hu T., Wang Y., Ye X., Zhan M., Ji C., Xu Z., Wang X., Gu Y, & Jia L. (2020). Induction of NEDD8-conjugating enzyme E2 UBE2F by platinum protects lung cancer cells from apoptosis and confers to platinum-insensitivity. Cell Death & Disease, 11(11), 975.
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3

Quantification of Apoptosis-Related Proteins

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Cells were homogenized in RIPA buffer, containing phosphatase and protease inhibitors (Beyotime). The protein from cell lysate (40 μg) was processed with SDS-PAGE and transferred to nitrocellulose paper. Then the following proteins were detected with appropriate antibodies: Bim (Cell Signaling), Ube2m (Abcam), Ube2f (Proteintech), Cullin1 (Abcam), Caspase 3 (Cell Signaling Technology), Cleaved-Caspase 3 (Cell Signaling Technology) and β-actin (Sigma-Aldrich). The band intensity was quantified with Image J software (NIH).
Zhang Y., Du L., Wang C., Jiang Z., Duan Q., Li Y., Xie Z., He Z., Sun Y., Huang L., Lu L, & Wen C. (2024). Neddylation is a novel therapeutic target for lupus by regulating double negative T cell homeostasis. Signal Transduction and Targeted Therapy, 9, 18.
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4

Quantifying Protein Expression in Cells and Extracellular Vesicles

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For western blotting (WB) assay, total cells and EVs were washed with ice‐cold PBS and lysed in SDS buffer on ice and boiled for 10 min at 100°C. Then, the samples were resolved by SDS‐PAGE followed by transfer onto PVDF membranes (Millipore) and probed with the indicated primary antibodies (CD63, Abclonal; Alix, Proteintech; Tsg101, Abclonal; CD81, Affinity; Coro1a, Abcam; GM130, Abcam; NEDD8, Abcam; Ub, CST; UBA3, Abcam; Cullin3, Sangon Biotech; UBE2F, Bioss; UBE2M, Abcam; TRIM4, Abclonal; Monla, Abclonal; Monlb, Abclonal; Rab7, Abcam; β‐Actin, Abclonal) and second antibodies (Goat anti‐mouse IgG HRP, Goat anti‐rabbit IgG HRP, MultiSciences).
For immunoprecipitation (IP) assays, cells were lysed in Co‐IP lysis buffer [50 mM Tris‐HCl, 5 mM EDTA, 150 mM NaCl, 0.5% (v/v) Nonidet‐P40, and 10% (v/v) glycerol (pH 7.4), supplemented with 1 mM PMSF, 1 mM Na3VO4, and 10 mM NaF]. The lysate was incubated with M2‐Flag beads (Sigma), anti‐His beads (MBL), anti‐HA beads (MBL) or anti‐Myc beads (Thermo Scientific) overnight at 4°C. The immunoprecipitates were washed at least three times in lysis buffer and then analysed by WB with the indicated antibodies (Flag, CST; His, MBL; HA, CST; Myc, Abmart).
Fei X., Li Z., Yang D., Kong X., Lu X., Shen Y., Li X., Xie S., Wang J., Zhao Y., Sun Y., Zhang J., Ye Z., Wang J, & Cai Z. (2021). Neddylation of Coro1a determines the fate of multivesicular bodies and biogenesis of extracellular vesicles. Journal of Extracellular Vesicles, 10(12), e12153.
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