Rabbit anti usp7

Manufactured by Fortis Life Sciences

Rabbit anti-USP7 is a primary antibody that specifically recognizes the Ubiquitin Specific Peptidase 7 (USP7) protein. USP7 is a deubiquitinating enzyme that plays a role in various cellular processes. This antibody can be used for the detection and analysis of USP7 in research applications.

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Anti-USP7 (7 citations)

4 protocols using rabbit anti usp7

Detailed Western Blot Protocol with Antibody List

Cited 2 times

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Western blots were performed as described previously^{82 (link),169 (link)}. The following antibodies were used:
Rabbit anti-UHRF1 (polyclonal; 1:250; Cell Signalling, D6G8E), mouse anti-alpha-Tubulin (monoclonal; 1:500; Sigma, T9026), rabbit anti-H3 (polyclonal; 1:1000; Abcam, ab1791), mouse anti-GFP (monoclonal; 1:1000; Roche), mouse anti-FLAG M2 (monoclonal; 1:1000; Sigma, F3165), rabbit anti-xDNMT1 (polyclonal;^{82 (link)}), rabbit anti-xUHRF1 (polyclonal;^{82 (link)}), rabbit anti-USP7 (polyclonal; Bethyl Lab., A300-033A), rabbit anti-H3 (polyclonal; Abcam, ab1791), rat anti-TET1 (monoclonal; 1:10;^{176 (link)}), rat anti-alpha-Tubulin (monoclonal; 1:250; Abcam, ab6160). goat anti-rat HRP (polyclonal; 1:1000; Jackson ImmunoResearch), goat anti-rabbit HRP (polyclonal; 1:1000; BioRad), mouse anti-xCDC45 (monoclonal;¹⁷⁷), mouse anti-xRPA2 (monoclonal;^{178 (link)}), and mouse anti-PCNA (monoclonal; Santa Cruz, sc56). Uncropped and unprocessed scans of blots can be found in the Source Data file.

Mulholland C.B., Nishiyama A., Ryan J., Nakamura R., Yiğit M., Glück I.M., Trummer C., Qin W., Bartoschek M.D., Traube F.R., Parsa E., Ugur E., Modic M., Acharya A., Stolz P., Ziegenhain C., Wierer M., Enard W., Carell T., Lamb D.C., Takeda H., Nakanashi M., Bultmann S, & Leonhardt H. (2020). Recent evolution of a TET-controlled and DPPA3/STELLA-driven pathway of passive DNA demethylation in mammals. Nature Communications, 11, 5972.

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Antibody-based Western Blotting and Immunoprecipitation

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The following antibodies were used for Western blotting or immunoprecipitation: anti-SF3B1 (Bethyl Laboratories, A300-996A-T, MBL International, D221-3), rabbit anti-cleaved Notch1 (Cell Signaling Technology, 4147S), rabbit anti–β-actin (Cell Signaling Technology, 4967S), rabbit anti-USP7 (Bethyl Laboratories, A300-033A), rabbit anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (Cell Signaling Technology, 2118S), rabbit anti-CHEK2 (Bethyl Laboratories, A300-618A), mouse immunoglobulin G2b (IgG2b) (MBL International, M077-3), P5091 (Selleck Chemicals, S7132), PR619 (Sigma-Aldrich, SML0430), b-AP15 (Selleck Chemicals, S4920), ML323 (Sigma-Aldrich, SML1177), BML277 (MedChem Express, HY-13946), BML277 (Sigma-Aldrich, C3742, in vivo), SCH900776 (MedChem Express, HY-15532), camptothecin (Selleck Chemicals, S1288), flavopiridol (Sigma-Aldrich, F3055), topotecan (Sigma-Aldrich, T2705), etoposide (Sigma-Aldrich, E1383), mitoxantrone (European pharmacopoeia reference standard, M2305000), doxorubicin (Sigma-Aldrich, 44583), and E7107 and H3B-8800 (H3 Biomedicine).

Han C., Khodadadi-Jamayran A., Lorch A.H., Jin Q., Serafin V., Zhu P., Politanska Y., Sun L., Gutierrez-Diaz B.T., Pryzhkova M.V., Abdala-Valencia H., Bartom E.T., Buldini B., Basso G., Velu S.E., Sarma K., Mattamana B.B., Cho B.K., Obeng R.C., Goo Y.A., Jordan P.W., Tsirigos A., Zhou Y, & Ntziachristos P. (2022). SF3B1 homeostasis is critical for survival and therapeutic response in T cell leukemia. Science Advances, 8(3), eabj8357.

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Western Blot Analysis of Ubiquitin Regulators

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A total of 10–50 μg protein was extracted from cells and used for Western blot analysis. Briefly, the cells were lysed using protein lysis buffer (50 mM Tris–Cl, 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 200 mM Na3VO4, 1x proteinase inhibitor, pH 7.4) and the protein concentration was measured using a Micro BCA^TM Protein Assay kit (Thermo Scientific), based on the standard curve of BSA. The antibodies used for immunoblot analysis were purchased and assessed for equal loading as follows: rabbit anti-USP7 (#A300-034, dilution ratio 1:1000) and rabbit anti-USP47 (#A301-048A, 1:1000) from Bethyl Laboratories; mouse anti-Flag (#F1804, 1:1000) from Sigma Aldrich; rabbit anti-ubiquitin (#3933, 1:2000) from Cell Signaling; mouse anti-p53 (#sc-126, 1:1000), and mouse anti-HSP90α/β (#sc-13119, 1:5000) from Santa Cruz Biotechnology; rabbit anti-β-actin (#LF-PA0207, 1:5000) from AbFrontier. Samples were analyzed using SDS-PAGE, and chemiluminescence was measured using the Ez-Capture MG imaging system (ATTO Corporation).

Shin S.C., Park J., Kim K.H., Yoon J.M., Cho J., Ha B.H., Oh Y., Choo H., Song E.J, & Kim E.E. (2023). Structural and functional characterization of USP47 reveals a hot spot for inhibitor design. Communications Biology, 6, 970.

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Western Blot Protein Detection Protocol

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Cells were lysed in 9 M urea, 10 mM Tris-HCl pH 6.8, sonicated and clarified by centrifugation. 80 µg of protein was subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were blocked in 5% non-fat dry milk in TBS-T (TBS with 0.1% Tween), then incubated with primary antibodies: mouse FLAG M2 (F1804 from Sigma; 1:5000 dilution), rabbit anti-FLAG (PA1-984B from Invitrogen; 1:5000 dilution), rabbit anti-HA (Santa Cruz 805; 1:200 dilution), rabbit anti-myc (Santa Cruz 789; 1:5000 dilution), rabbit anti-USP7 (Bethyl laboratories A300-033A; 1:10000 dilution), mouse anti-KIF20B (Santa Cruz 515194; 1:200 dilution), rabbit anti-FBXO38 (Bethyl laboratories A302-378A; 1:5000 dilution), rabbit anti-SKP1 (Santa Cruz 7163; 1:200 dilution), goat anti-actin (Santa Cruz 1615; 1:1000 dilution). Membranes were washed three times in TBS-T, followed by incubation with goat anti-mouse HRP (Santa Cruz 2005), goat anti-rabbit HRP (Sigma SAB3700878) or donkey anti-goat HRP (Sigma SAB3700285) at 1:5000 dilution. Membranes were developed using chemiluminescence reagents (ECL or ECL-prime; Santa Cruz or Amersham). Protein bands were quantified using ImageQuantTL (GE Healthcare Sciences) and normalized to actin bands.

Georges A., Coyaud E., Marcon E., Greenblatt J., Raught B, & Frappier L. (2019). USP7 Regulates Cytokinesis through FBXO38 and KIF20B. Scientific Reports, 9, 2724.

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