UbcH5C (D60E2) Rabbit mAb 4330S from Cell Signaling Technology

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International journal of molecular sciences
January 20 2022

Fig 1: Effects of the sesquiterpene lactone, BdS, on UBE2D activity in vitro and in cells. (A) COBALT Alignment and ESPript visualisation of UniProt sequences of UBE2D1-4 [15,16]. Red shading of single-letter amino acid codes indicates homology; white shading shows points of differentiation between enzymes. Red arrow points out catalytic cysteine, C85. Secondary structure elements are displayed at the top for UBE2D1. UBE2D1 accessibility is indicated at the bottom (colour gradient—darker blue to white showing decreasing accessibility). (B) Overlay of available UBE2D crystal structures (UBE2D1: PDB 5TUT (red), UBE2D2: PDB 2ESK (green), UBE2D3: PDB 3L1Z (blue)) [17,18,19]. Red arrow indicates catalytic cysteine. (C) Chemical structures of sesquiterpene lactones, ATL and IJ-5 (alantolactone, and 1ß-hydroxyalantolactone, respectively), and BdS showing their shared covalent-binding warhead in blue. (D) Left: In vitro ubiquitylation assay workflow (created using Biorender). Right: Representative blots showing the effects of BdS on ubiquitin-loading for UBE2D1 (n = 2 independent experiments) and UBE2D3 (n = 3 independent experiments). (E) Doxycycline (Dox) induction of GFP-UBE2D1 wt (wild-type) and GFP-UBE2D1 CD (catalytically dead—C85S; n = 1 experiment) in U2OS cells. (F) Fluorescence images of GFP-UBE2D1 wt and GFP-UBE2D1 CD in siCTRL or siALL-Ds depleted U2OS cells, after doxycycline (Dox) induction or not. Nuclei are outlined in white using DAPI as a reference (scale bar – 20 µm). (G) GFP-Trap pulldowns of GFP-UBE2D1 stably expressed in U2OS cells. IP represents 1% of the input (n = 2 independent experiments). Immunoblot sections are derived from same membrane. (H) siALL-Ds depletion efficiency of endogenous (endo.) UBE2D1 (D1) in U2OS cells stably expressing GFP. Blot is representative of n = 4 independent experiments. (I) Effects of PYR-41 (left), a ubiquitin E1 (UBA1) inhibitor, and BdS (right) on GFP-UBE2D1 wt auto-ubiquitylation at the indicated concentrations and treatment times (n = 1 for PYR-41; n = 2 independent experiments for BdS). Normalised blot band intensity quantification was performed using ImageJ.
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Molecular Cancer
December 01 2022

Fig 2: UbcH5c knockdown decreases the anticancer activity of DHPO. a and b The expression levels of indicated proteins in Panc1 and SW1990 cells transfected with shRNA targeting UbcH5c or control shRNA were determined by western blotting analysis after treatment with DHPO for 24 h. c and d The 72-h cell viability assays in indicated cell lines transfected with shRNA targeting UbcH5c or control shRNA. e Scheme showing a proposed molecular mechanism for the anticancer activity of DHPO
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Nature communications
June 11 2018

Fig 3: SKP2 interacts with YAP and induces its K63-linked polyubiquitination. a The HEK293T SFB-YAP stable cell line was transfected with MYC-tagged E3 ligases, followed by immunoprecipitation with anti-MYC beads and immunoblotting with antibodies against FLAG and MYC. b The HEK293T SFB-YAP stable cell line was co-transfected with the K63-specific mutant of HA-ubiquitin and the indicated E3 ligase, followed by pulldown with S-protein beads and immunoblotting with antibodies against HA and FLAG. c Co-immunoprecipitation of endogenous SKP2 with endogenous YAP. d In vitro binding of bacterially purified MBP-SKP2 to in vitro translated and purified GST-YAP. e In vitro binding of bacterially purified MBP-SKP2 to SFB-YAP purified from the HEK293T SFB-YAP stable cell line. MBP-SKP2 or MBP protein was incubated with SFB-YAP protein with or without CIP treatment at 37 °C for 1 h, followed by immunoblotting with antibodies against SKP2, p-YAP (S127), and FLAG. f The HEK293T SFB-YAP stable cell line was transfected with HA-ubiquitin (wild-type, K48R or K63R) and MYC-SKP2, followed by pulldown with S-protein beads and immunoblotting with antibodies against HA and FLAG. g The HEK293T SFB-YAP stable cell line was transfected with HA-ubiquitin (wild-type, K48-specific or K63-specific) and MYC-SKP2, followed by pulldown with S-protein beads and immunoblotting with antibodies against HA and FLAG. h Total K63-linkage specific ubiquitinated proteins in control or SKP2 knockout HEK293A cells (generated by CRISPR-Cas9) were immunoprecipitated by a K63-linked polyubiquitin-specific antibody, followed by immunoblotting with antibodies against YAP and ubiquitin. i siRNA targeting CUL1 or SKP1 was transfected into the HEK293T SFB-YAP stable cell line. Forty-eight hours after siRNA transfection, cells were transfected with HA-ubiquitin and MYC-SKP2, followed by pulldown with S-protein beads and immunoblotting with antibodies against HA and FLAG. j The HEK293T UBCH5c shRNA stable cell line was transfected with SFB-YAP and HA-ubiquitin with or without MYC-SKP2, followed by pulldown with S-protein beads and immunoblotting with antibodies against HA and FLAG. k Purified GST-YAP protein was incubated with ATP, E1, UBCH5c, and the K63-specific mutant of His-ubiquitin with or without the SCFSKP2 complex, followed by immunoprecipitation with a YAP-specific antibody and immunoblotting with antibodies against ubiquitin and YAP
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Molecular Cancer
December 01 2022

Fig 4: UbcH5c (also named UBE2D3) is overexpressed in pancreatic cancer and correlated with the poor prognosis of patients with this disease. a Heatmap showing mRNA expression level correlation of E2s with IAP family members, including cIAP1/BIRC2, cIAP2/BIRC3, and XIAP. b Kaplan–Meier survival analysis for the association of UbcH5c expression with overall survival of patients in the TCGA pancreatic cancer dataset. c UbcH5c mRNA expression is upregulated in the pancreatic tumor tissues compared with it in the normal tissues group from TCGA TARGET GTEx dataset. d UbcH5c mRNA expression is upregulated in the pancreatic tumor samples compared with paired adjacent nontumorous samples from TCGA dataset. e Representative IHC images on tissue microarray probed with the anti-UbcH5c antibody in Human Protein Atlas. f Correlation analysis of UbcH5c and indicated genes mRNA expression levels in the TCGA pancreatic cancer dataset. Correlation coefficient r and P values are shown on the graphs. * P < 0.05, ** P < 0.01, *** P < 0.001
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Molecular Cancer
December 01 2022

Fig 5: DHPO directly targets UbcH5c. a, b and c The predicted binding mode of DHPO to UbcH5c by docking analysis. d Chemical structure of DHPO. e and f The binding affinity of DHPO to UbcH5c was evaluated by surface plasmon resonance analysis. g The DHPO-UbcH5c binding was examined in Panc1 and SW1990 cells using cellular thermal shift assays and western blot

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Cell Signaling Technology for
UbcH5C (D60E2) Rabbit mAb

Cell Signaling Technology's UbcH5C (D60E2) Rabbit mAb is a Rabbit monoclonal antibody. The UbcH5C (D60E2) Rabbit mAb Antibody was generated using UBE2D3 as the antigen. It reacts with Homo Sapiens, Human, Mouse, Rat, and Non-Human Primate. This antibody has been shown to work in applications such as: Precipitation, Immunoprecipitation, and Western Blot.

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UbcH5C (D60E2) Rabbit mAb detects endogenous levels of total UbcH5C protein

Product Specs
ItemUbcH5C (D60E2) Rabbit mAb
CompanyCell Signaling Technology
Price Pricing Info Supplier Page View Company Product Page
Catalog Number4330S
Quantity100 µl
ApplicationsW, IP
ReactivityH, M, R, Mk
TargetUBCH5C
HostRabbit
NCBI Gene AliasesE2(17)KB3, UBC4/5, UBCH5C
ClonalityMonoclonal
NCBI Full Gene Nameubiquitin conjugating enzyme E2 D3
Research AreaUbiquitin and Ubiquitin-like proteins
IsotypeIgG
CloneD60E2
Molecular Weight14

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(1) Keratoconus Diagnosis: From Fundamentals to Artificial Intelligence: A Systematic Narrative ReviewDiagnostics (Basel)August 21, 2023Sana Niazi, Marta Jiménez-García, Oliver Findl, Zisis Gatzioufas, Farideh Doroodgar, Mohammad Hasan Shahriari, Mohammad Ali JavadiCorvis ST (CST; Oculus, Wetzlar, Germany) is a non-contact tonometer that uses a Scheimpflug camera with 4330 frames per second to record the ocular response to an air pulse in an 8 mm wide horizontal section of

(2) Corneal biomechanics after small incision lenticule extraction and femtosecond laser in situ keratomileusis: A systematic review and meta-analysisMDAugust 11, 2023Songbai Chen, Hongjie Ma, Congling Zhaocorneal deformation in response to air pulses within 100 ms using an ultra-fast Scheimpflug camera (4330 frames/seconds). Unlike ORA, CST can record corneal deformation data in real time to analyze corneal

(3) Comparison of Corvis ST Parameters between Primary Open-Angle Glaucoma and Primary Angle-Closure GlaucomaJ Clin MedAugust 3, 2023Yuta Nakaniida, Kana Tokumo, Hiromitsu Onoe, Naoki Okada, Shunsuke Nakakura, Ryo Asaoka, Yoshiaki Kiuchiimages that captured corneal deformation following a rapid air puff. The device was capable of capturing 4330 images per second, which were analyzed to quantify CCT, applanation time, applanation length, corneal

(4) UHRF1 is essential for proper cytoplasmic architecture and function of mouse oocytes and derived embryosLife science allianceAugust 1, 2023Uemura S, Maenohara S, Inoue K, Ogonuki N, Matoba S, Ogura A, Kurumizaka M, Yamagata K, Sharif J, Koseki H, et al.milk and incubated with primary antibodies against FBXO38 (1:1,000, ab87729; Abcam), UBE2D3 (1:1,000, 4330; Cell Signaling Technology), or SPIN1 (1:1,000, 12105-1-AP; Proteintech). After washing several times,

(5) Ubiquitinome Profiling Reveals in Vivo UBE2D3 Targets and Implicates UBE2D3 in Protein Quality ControlMolecular & Cellular ProteomicsJune 1, 2023Zeliha Yalçin, Daniëlle Koot, Karel Bezstarosti, Daniel Salas-Lloret, Onno B. Bleijerveld, Vera Boersma, Mattia Falcone, Román González-Prieto, Maarten Altelaar, Jeroen A.A. Demmers, et al.Thermo Scientific). Primary antibodies used were against UBE2D (A615, Boston Biochem, 1:2000 dilution or 4330S, CST, 1:500 dilution), V5 (R960-25, Invitrogen, 1:1000 dilution), HA (MMS-101R, Covance, 1:1000 dilution),

(6) A bio-orthogonal linear ubiquitin probe identifies STAT3 as a direct substrate of OTULIN in glioblastomaNucleic acids researchFebruary 22, 2023Du X, Pang J, Gu B, Si T, Chang Y, Li T, Wu M, Wang Z, Wang Y, Feng J, et al.CST), human anti-linear Ub (14) (a gift from Dr Vishva M. Dixit, Genentech Inc.), rabbit anti-UBE2D3 (4330S, CST), rabbit anti-OTULIN (ab15117, Abcam), mouse anti-α-tubulin (T5168, Sigma), mouse anti-OLIG2 (66513,

(7) Targeting E2 ubiquitin-conjugating enzyme UbcH5c by small molecule inhibitor suppresses pancreatic cancer growth and metastasisMolecular CancerDecember 1, 2022Simin Qi, Xiaoqing Guan, Jia Zhang, Dehua Yu, Xuefei Yu, Qinglin Li, Wenjuan Yin, Xiang-Dong Cheng, Weidong Zhang, Jiang-Jiang Qinanti-c-Myc (Cat. No. 18583S, 1/1000), anti-Mcl-1 (Cat. No. 94296S, 1/1000), and anti-UbcH5c (Cat. No. 4330S, 1/1000) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). In vitro assays

(8) Dynamic quality control machinery that operates across compartmental borders mediates the degradation of mammalian nuclear membrane proteinsCell ReportsNovember 22, 2022Pei-Ling Tsai, Christopher J.F. Cameron, Maria Fernanda Forni, Renee R. Wasko, Brigitte S. Naughton, Valerie Horsley, Mark B. Gerstein, Christian Schlieker14773S; RRID: AB_2798607 Rabbit monoclonal anti-UbcH5C clone D60E2,Cell Signaling Technology,Cat# 4330S; RRID: AB_10544697 Mouse monoclonal anti-Lamin A/C clone 4C11,Cell Signaling Technology,Cat# 4777S;

(9) Interactomic analysis reveals a homeostatic role for the HIV restriction factor TRIM5α in mitophagyCell reportsMay 10, 2022Saha B, Salemi M, Williams GL, Oh S, Paffett ML, Phinney B, Mandell MA.,Cell Signaling Technology,#3504; RRID: AB_2255663 UbcH5C (D60E2) Rabbit mAb,Cell Signaling Technology,#4330; RRID: AB_10544697 UBE2N (D2A1) Rabbit mAb,Cell Signaling Technology,#6999; RRID: AB_10828936 Atg13 (E1Y9V) Rabbit mAb

(10) Sesquiterpene Lactones Potentiate Olaparib-Induced DNA Damage in p53 Wildtype Cancer CellsInternational journal of molecular sciencesJanuary 20, 2022Osborne HC, Larrosa I, Schmidt CK.cat#ab176561—1:10,000 in 1% BSA in TBST 1×), and anti-UBE2D3 (Cell Signalling Technology; Danvers, MA, USA; cat#4330, RRID: AB_10544697—1:1000 in 1% BSA in TBST 1×). The following secondary antibodies were used for immunoblotting

(11) Identification of microRNAs that promote erlotinib resistance in non-small cell lung cancerBiochemical PharmacologyJuly 1, 2021A.S. Pal, M. Bains, A. Agredo, A.L. Kasinskicells hsa-miR-5693,4.806839,hsa-miR-5693,6.980146 hsa-miR-3618,4.128858,hsa-miR-432-5p,5.680975 hsa-miR-3657,3.53778,hsa-miR-3618,5.201033 hsa-miR-432-5p,3.447473,hsa-miR-4787-5p,3.098385 hsa-miR-4435,3.369496,hsa-miR-3198,2.666229 hsa-miR-588,3.115648,hsa-miR-4435,2.599998 hsa-miR-4701-3p,3.096875,hsa-miR-588,2.331268 hsa-miR-4787-5p,2.550594,hsa-miR-4693-5p,2.305733 hsa-miR-4521,2.406701,hsa-miR-4797-5p,2.231674 hsa-miR-4499,2.176414,hsa-miR-4775,2.208845 hsa-miR-9-5p,2.139334,hsa-miR-4521,2.115477 hsa-miR-4643,2.074379,hsa-miR-4643,2.072277 hsa-miR-4486,1.758585,hsa-miR-4781-3p,1.938728 hsa-miR-548ac,1.743734,hsa-miR-4757-5p,1.797722 hsa-miR-4757-5p,1.71694,hsa-miR-4328,1.77053 hsa-miR-3926,1.614334,hsa-miR-4638-3p,1.730049 hsa-miR-4760-5p,1.552469,hsa-miR-4329,1.706815 hsa-miR-4748,1.508298,hsa-miR-4634,1.588003 hsa-miR-4792,1.494049,hsa-miR-4499,1.475899 hsa-miR-3198,1.474083,hsa-miR-4791,1.467733 hsa-miR-5694,1.472514,hsa-miR-4792,1.452575 hsa-miR-4778-5p,1.463761,hsa-miR-3665,1.405999 hsa-miR-4638-3p,1.358205,hsa-miR-4701-3p,1.387164 hsa-miR-3677-5p,1.339528,hsa-miR-4753-3p,1.364724 hsa-miR-4794,1.313135,hsa-miR-4793-5p,1.352131 hsa-miR-4693-5p,1.313076,hsa-miR-4706,1.338108 hsa-miR-4797-5p,1.298536,hsa-miR-4698,1.306059 hsa-miR-4328,1.276663,hsa-miR-4488,1.281768 hsa-miR-4753-3p,1.207611,hsa-miR-4778-5p,1.251342 hsa-miR-4781-3p,1.206249,hsa-miR-4740-3p,1.224541 hsa-miR-4488,1.182468,hsa-miR-4330,1.168158 hsa-miR-4740-3p,1.181966,hsa-miR-4748,1.116061 hsa-miR-548ab,1.140096,hsa-miR-3677-5p,1.110286 hsa-miR-4793-5p,1.126522,hsa-miR-4794,1.103088 hsa-miR-4634,1.121035,hsa-miR-4705,1.09914 hsa-miR-4705,1.119791,hsa-miR-4760-5p,1.035345 hsa-miR-4791,1.107927,hsa-miR-4728-5p,0.980355 hsa-miR-4706,1.104596,hsa-miR-548ak,0.893768 hsa-miR-4522,1.043842,hsa-miR-3926,0.810352 hsa-miR-4775,1.017418,hsa-miR-5694,0.804238 hsa-miR-548ak,0.971604,hsa-miR-9-5p,0.732981 hsa-miR-4468,0.943862,hsa-miR-548ac,0.313635 hsa-miR-3665,0.934691,hsa-miR-548ab,0.232357 hsa-miR-548aj-3p,0.926283,hsa-miR-4676-3p,0.222049 hsa-miR-4751,0.891378,hsa-miR-4654,0.120642 hsa-miR-4330,0.883232,hsa-miR-4661-3p,0.244 hsa-miR-4654,0.851533,hsa-miR-4741,0.065 hsa-miR-4661-3p,0.844637,hsa-miR-4516,−0.189 hsa-miR-4329,0.841927,hsa-miR-4751,0.040 hsa-miR-4698,0.837794,hsa-miR-204-3p,−0.128 hsa-miR-4799-3p,0.821699,hsa-miR-4690-3p,−0.318 hsa-miR-4728-5p,0.798065,hsa-miR-4486,−0.225 hsa-miR-4534,0.788234,hsa-miR-548ad,−0.360 hsa-miR-548ad,0.7357,hsa-miR-4468,−0.590 hsa-miR-4762-3p,0.661747,hsa-miR-4522,0.797 hsa-miR-4516,0.544357,hsa-miR-4762-3p,−0.248 hsa-miR-4741,0.529365,hsa-miR-3657,−0.908 hsa-miR-204-3p,0.424363,hsa-miR-548aj-3p,−0.128 hsa-miR-4690-3p,0.383958,hsa-miR-4799-3p,−1.572 hsa-miR-4676-3p,0.331065,hsa-miR-4534,−1.743 The

(12) Corneal Distortion and Corneal Morphological Parameters in NaÃ¯ve Eyes Analyzed with a Scheimpflug CameraModern Health ScienceDecember 30, 2020Sandro Sbrodone, Adele Ragucci, Gennarfrancesco Iaccarino, Domenico de Robertis, Giovan Battista Scazzi, Michele Lanzathe corneal behavior during an air puff indentation thanks to a Scheimpflug camera able to process 4330 images per second, and it shows many corneal deformation parameters that are well defined (Figure 1)

(13) Intraocular pressure measurement techniques: Current concepts and a reviewIndian Journal of Clinical and Experimental OphthalmologySeptember 28, 2020Hemant Sharma, Sanjeev K Nainiwal, Akshay Sarraf, Rakesh Porwal, Vijaya Sharmaan ultrahigh-speed Scheimpflug camera equipped to record the movements of the cornea. The CST gathers 4330 frames per second within a 100- millisecond period, therefore recording dynamic deformation of the cornea

(14) Lysosomes, Autophagy, and Hormesis in Cell Physiology, Pathology, and Age-Related DiseaseDose-ResponseJuly 7, 2020Michael N. Moores41559-017-0116 Mijaljica D, Devenish RJ. Nucleophagy at a glance. J Cell Sci. 2013;126(Pt 19):4325-4330. De P, Miskimins K, Dey N, et al. Promise of rapalogues versus mTOR kinase inhibitors in subset specific

(15) Correlation Between the Myopic Retinal Deformation and Corneal Biomechanical Characteristics Measured With the Corvis ST TonometryTranslational Vision Science & TechnologyAugust 19, 2019Shotaro Asano, Ryo Asaoka, Takehiro Yamashita, Shuichiro Aoki, Masato Matsuura, Yuri Fujino, Hiroshi Murata, Shunsuke Nakakura, Yoshitaka Nakao, Yoshiaki Kiuchipuff pulse during the inward and outward movements with a high-speed Scheimpflug camera, capturing 4330 frames per second. This imaging system allows us to investigate the dynamic inspection of the actual

(16) SKP2- and OTUD1-regulated non-proteolytic ubiquitination of YAP promotes YAP nuclear localization and activityNature communicationsJune 11, 2018Yao F, Zhou Z, Kim J, Hang Q, Xiao Z, Ton BN, Chang L, Liu N, Zeng L, Wang W, et al.Scientific). Antibodies against YAP (14074, 1:1000), phospho-YAP (4911, 1:500), SKP2 (2652, 1:1000), UBCH5c (4330, 1:500), GST (2624, 1:1000), Angiomotin (43130, 1:1000), TEAD (13295, 1:500), 14-3-3 (8312, 1:500),

(17) Association between Corneal Biomechanical Properties with Ocular Response Analyzer and Also CorvisST Tonometry, and Glaucomatous Visual Field SeverityTranslational Vision Science & TechnologyJune 14, 2017Kazunori Hirasawa, Masato Matsuura, Hiroshi Murata, Shunsuke Nakakura, Yoshitaka Nakao, Yoshiaki Kiuchi, Ryo Asaokadetail elsewhere.26 In short, the instrument's camera records a sequence of images (capable of capturing 4330 images per second) that capture corneal deformation due to the application of a rapid air puff (Fig.).

(18) Using CorvisST tonometry to assess glaucoma progressionPLOS ONEMay 4, 2017Masato Matsuura, Kazunori Hirasawa, Hiroshi Murata, Shunsuke Nakakura, Yoshiaki Kiuchi, Ryo Asaokabetween each measurement. As described in detail elsewhere,[26] CST captures a sequence of images (4330 images per second) of corneal deformation, and various parameters of deformation amplitude, applanation

(19) Cataract surgery causes biomechanical alterations to the eye detectable by Corvis ST tonometryPLOS ONEFebruary 21, 2017Yoshitake Kato, Shunsuke Nakakura, Ryo Asaoka, Kanae Matsuya, Yuki Fujio, Yoshiaki Kiuchi, on behalf of the Ocular Biomechanics Study Grouptonometer (Corvis ST tonometry: CST, Oculus, Wetzlar, Germany) is equipped with a high-speed camera (4330 frames per second) that can capture a series of horizontal Scheimpflug images during corneal deformation

(20) Summertime ozone formation in Xi'an and surrounding areas, ChinaAtmospheric Chemistry and PhysicsApril 7, 2016Tian Feng, Naifang Bei, Ru-Jin Huang, Junji Cao, Qiang Zhang, Weijian Zhou, Xuexi Tie, Suixin Liu, Ting Zhang, Xiaoli Su, et al.predicting the wind speed and direction at the Chang’an and Lantian Atmos. Chem. Phys., 16, 4323–4342, 2016 4330 T. Feng et al.: Summertime ozone formation in Xi’an and surrounding areas Figure 8. Pattern comparison

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International journal of molecular sciences

Molecular Cancer

Nature communications

Molecular Cancer

Molecular Cancer

UbcH5C (D60E2) Rabbit mAb from Cell Signaling Technology

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Cell Signaling Technology

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UbcH5C (D60E2) Rabbit mAb from Cell Signaling Technology

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