Anti usp14

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-USP14 is a laboratory reagent that targets the protein USP14, which is involved in the ubiquitin-proteasome system. It can be used for research purposes to study the role and function of USP14 in various cellular processes.

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Lab products found in correlation

Anti actin, by Merck Group (2 mentions) Anti β catenin, by Santa Cruz Biotechnology (2 mentions) Anti ub, by Santa Cruz Biotechnology (2 mentions) Propidium iodide, by Merck Group (1 mentions) N acetylcysteine, by Merck Group (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Anti grp78, by Santa Cruz Biotechnology (1 mentions) Anti uchl5, by Abcam (1 mentions) Anti cofilin 1, by Santa Cruz Biotechnology (1 mentions) Anti pcofilin 1, by Santa Cruz Biotechnology (1 mentions) Bortezomib, by LC Laboratories (1 mentions) Anti myc, by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions) Anti usp14, by Cell Signaling Technology (1 mentions) Anti beclin 1, by Santa Cruz Biotechnology (1 mentions) Mg132 s2619, by Selleck Chemicals (1 mentions) Anti ha, by Proteintech (1 mentions) Anti myc, by Proteintech (1 mentions) Anti phospho akt ser473, by Cell Signaling Technology (1 mentions) Anti tubulin, by Proteintech (1 mentions)

6 protocols using anti usp14

Bortezomib-Induced Apoptosis Pathway

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Chemicals used were: bortezomib (LC Laboratories), Propidium Iodide and N-acetylcysteine (Sigma Aldrich). Primary antibodies used were: anti-Gas2^{36 (link)} anti-actin (Sigma/Aldrich) anti-USP1^{37 (link)} anti-USP33 (Millipore) anti-AKT (Cell Signaling) anti-UCH-L5 (Abcam), anti-USP18^{38 (link)} anti-HDAC4^{39 (link)} anti-MEF2D (BD Bioscience), anti-GRP78, anti-USP14, anti-Cofilin-1, anti-pCofilin-1 (Santa Cruz Biotechnologies).

Ciotti S., Sgarra R., Sgorbissa A., Penzo C., Tomasella A., Casarsa F., Benedetti F., Berti F., Manfioletti G, & Brancolini C. (2018). The binding landscape of a partially-selective isopeptidase inhibitor with potent pro-death activity, based on the bis(arylidene)cyclohexanone scaffold. Cell Death & Disease, 9(2), 184.

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Western Blot Antibody Panel for Autophagy

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The commercial antibodies used for Western blot analysis include the following: Anti-phospho-Akt (Ser473) (1:1000 dilution; no. 3787), anti-LC3B (1:1000 dilution; no. 2775), anti-USP14 (rabbit, 1:1000 dilution; no. 11931), anti-Atg14L (1:1000 dilution; no. 5504), and anti-Flag (1:1000 dilution; no. 2368) were from Cell Signaling Technology. Anti-Beclin 1 (1:1000 dilution for Western blot [sc-11427]; 1:200 dilution for immunoprecipitation [sc-48341]), anti-Akt (1:1000 dilution; sc-8312), and anti-USP14 (mouse, 1:100 dilution for immunoprecipitation; sc-393872) were from Santa Cruz Biotechnology. Anti-Myc (1:1000 dilution; 16286-1-AP), anti-HA (1:1000 dilution; 51064-2-AP) anti-Rubion (1:1000 dilution; 21444-1-AP), and anti-GAPDH (1:10,000 dilution; 60004-1-Ig) were from Proteintech. Anti-Tubulin (1:10,000 dilution; PM054), anti-UVRAG (1:1000 dilution; PD027), and anti-p62 (1:5000; PM045) were from MBL. Anti-ubiquitin Lys63-specific antibody (1:2000 dilution; no. 2210353) was from Milllipore. Mouse monoclonal anti-Flag (1:500 for immunoprecipitation; F1804), Anti-Myc, and anti-HA affinity gel (E6779) were from Sigma-Aldrich. IU1 (S7134), MK2206 (S1078), E64D (S7379), rapamycin (S1039), and MG132 (S2619) were from Selleckchem.

Xu D., Shan B., Sun H., Xiao J., Zhu K., Xie X., Li X., Liang W., Lu X., Qian L, & Yuan J. (2016). USP14 regulates autophagy by suppressing K63 ubiquitination of Beclin 1. Genes & Development, 30(15), 1718-1730.

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Molecular Interplay of Ubiquitin Proteases

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bAP15 (11324) was purchased from Cayman Chemical (Ann Arbor, MI). The USP14 inhibitor IU1 (662210) was obtained from EMD Millipore Corporation (Billerica, MA). UCHL5 shRNA plasmid (h) (sc-76797-SH) was purchased from Santa Cruz Biotechnology (Dallas, TX). HA-ubiquitin (Plasmid 18712) was purchased from Addgene (Cambridge, MA).
Antibodies were obtained as follows; anti-β-actin (sc-47778), anti-UCHL5 (sc-271002), anti-USP14 (sc-100630), anti-β-catenin (sc-7963), anti-Vimentin (sc-6260), anti-Ub (sc-166553), and anti-HA-probe (F-7) (sc-7392) (Santa Cruz Biotechnology), anti-E-cadherin (610181), anti-N-cadherin (610920)(BD Bioscience, San Jose, CA), anti-Smad2 (5339), anti-Phospho-Smad2 (3108), anti-Phospho-Smad2 (18338), anti-Smad3 (9523), anti-Phospho-Smad3 (9520), and anti-Smad4 (38454) (Cell Signaling Technology, Danvers, MA).

Fukui S., Nagasaka K., Miyagawa Y., Kikuchi-Koike R., Kawata Y., Kanda R., Ichinose T., Sugihara T., Hiraike H., Wada-Hiraike O., Sasajima Y, & Ayabe T. (2019). The proteasome deubiquitinase inhibitor bAP15 downregulates TGF-β/Smad signaling and induces apoptosis via UCHL5 inhibition in ovarian cancer. Oncotarget, 10(57), 5932-5948.

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Protein Expression Analysis in ESCC

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Total protein was extracted from ESCC tissues or cells using radioimmunoprecipitation assay (RIPA) lysis buffer containing a mixture of protease inhibitor. The protein concentration was quantified by the Bradford assay. A total of 50 μg of proteins was separated by 10% SDS–polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes (Amersham, Little Chalfont, UK). After blocking with 5% fat-free milk in TBS [20 mmol/L Tris, 0.15 mol/L NaCl (pH 7.0), 0.1% Tween 20], the membranes were incubated with primary antibodies (anti-USP14, anti-E-cadherin, anti-N-cadherin, anti-β-catenin, anti-cyclin D1, anti-c-Myc, and anti-GAPDH) (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C overnight. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology) for 1 h at room temperature. Finally, the blots were visualized by enhanced chemiluminescence (ECL) detection system (Amersham). The gray intensity analysis was performed using the Image-Pro Plus 6.0 software (Media Cybernetics Inc., Rockville, MD, USA).

Zhang J., Zhang D, & Sun L. (2017). Knockdown of Ubiquitin-Specific Protease 14 (USP14) Inhibits the Proliferation and Tumorigenesis in Esophageal Squamous Cell Carcinoma Cells. Oncology Research, 25(2), 249-257.

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Dissecting TRAIL-induced Apoptosis Signaling

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Sigma Chemical Co. provided honokiol, cycloheximide and MG132 (St. Louis, MO, USA), and R&D system supplied recombinant human recombinant TRAIL and z-VAD-fmk (Minneapolis, MN, USA). Enzo Life Sciences provided lactacystin (Ann Arbor, MI, USA). The primary antibodies were as follows: Cell Signaling Technology supplied anti-PARP, anti-cleaved caspase-3, anti-Bcl-xL, anti-DR5, anti-CHOP, and anti-UCHL1 (Beverly, MA, USA). Sigma Chemical Co. supplied anti-actin (St. Louis, MO, USA). Enzo Life Sciences provided anti-pro-caspase-3 and anti-c-FLIP (San Diego, CA, USA). BD Biosciences provided anti-Bim and anti-XIAP (San Jose, CA, USA). Abcam supplied anti-DR4 (Cambridge, MA, USA). R&D system supplied anti-survivin (Minneapolis, MN, USA). Santa Cruz Biotechnology provided anti-Mcl-1, anti-Bcl-2, anti-cIAP2, anti-ATF4, anti-Ub, anti-Cbl, anti-Itch, anti-USP14, anti-USP33, anti-OTUB1, anti-TRABID, and anti-STAMBPL1 (St. Louis, MO, USA). Bethyl Laboratories Inc provided anti-USP7 and anti-USP8 (Montgomery, TX, USA). Novus Biologicals supplied anti-USP53 (Centennial, CO, USA). Abnova provided anti-USP9X (Taipei City, Taiwan). The siRNAs were as follows: GFP (control) siRNA (Bioneer, Daejeon, Korea), DR5 siRNA (Invitrogen, Carlsbad, CA, USA), and STAMBPL1 siRNA (Santa Cruz Biotechnology, St. Louis, MO, USA). STAMBPL1 plasmid was a gift from Dr. H.C. Kang (Ajou University, Suwon, Korea).

Woo S.M., Seo S.U., Kubatka P., Min K.J, & Kwon T.K. (2019). Honokiol Enhances TRAIL-Mediated Apoptosis through STAMBPL1-Induced Survivin and c-FLIP Degradation. Biomolecules, 9(12), 838.

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Protein Analysis via Western Blotting and IP

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Western blotting and immunoprecipitation assays were performed as previously described (18) . Primary antibodies used in Western blotting are anti-KIF15 (1:1,000, catalog no. 55407-1-AP; Protein-Tech), anti-AR (1:1,000, catalog no. 06-680; Millipore; RayBiotech catalog no. 130-10083-1000, RRID:AB_11218813), anti-AR-V7 (1:1,000, catalog no. 19672; Cell Signaling Technology), anti-USP14 (1:1,000, catalog no. sc-393872; Santa Cruz Biotechnology), anti-Ubiquitin (1:1,000, catalog no. 3936; Cell Signaling Technology; LifeSpan catalog no. LS-C93201-1000, RRID:AB_10641875), anti-His (1:1,000, catalog no. 12698; Cell Signaling Technology; LifeSpan catalog no. LS-C129774-1000, RRID:AB_10832018), and anti-GAPDH (1:1,000, catalog no. ab181602; Abcam).

Gao L., Zhang W., Zhang J., Liu J., Sun F., Liu H., Hu J., Wang X., Wang X., Su P., Chen S., Qu S., Shi B., Xiong X., Chen W., Dong X, & Han B. (2021). KIF15-Mediated Stabilization of AR and AR-V7 Contributes to Enzalutamide Resistance in Prostate Cancer. Cancer research, 81(4).

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