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Anti ub sc 8017

Manufactured by Santa Cruz Biotechnology
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Anti-Ub (sc-8017) is a primary antibody product offered by Santa Cruz Biotechnology. It is directed against ubiquitin, a small regulatory protein found in eukaryotic cells. The antibody can be used to detect and study ubiquitin and its associated cellular processes.

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Lab products found in correlation

Anti flag f3165, by Merck Group (3 mentions) Anti v5 r960 25, by Thermo Fisher Scientific (2 mentions) Rabbit anti akt pt308, by Cell Signaling Technology (2 mentions) Anti ps473 akt, by Cell Signaling Technology (2 mentions) Anti nedd4l 13690 1 ap, by Proteintech (2 mentions) P s6s236 236, by Cell Signaling Technology (2 mentions) Protein assay reagent, by Bio-Rad (2 mentions) Anti pan akt, by Cell Signaling Technology (2 mentions) Anti s6, by Cell Signaling Technology (2 mentions) Phosphatases inhibitors, by Merck Group (2 mentions) Protease inhibitor, by Roche (2 mentions) Anti ha tag, by Cell Signaling Technology (2 mentions) Anti gm130 610822, by BD (1 mentions) 17 dmag, by Selleck Chemicals (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Merck Group (1 mentions) Anti acetylated α tubulin sc 23950, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488 or 546 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Antifade mounting medium, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Tunel fitc apoptosis detection kit, by Vazyme (1 mentions)

Close spelling variants of this product

Anti‐ub (sc‐8017) Abs Anti-YAP1 (SC15407) and anti-Ub (sc-8017) antibodies Anti-Ubiquitin (Ub) (sc-8017) Ub (sc-8017) Sc-8017 Anti-Ub

13 protocols using anti ub sc 8017

1

Antibodies for Hsp90, Tubulin, and Cell Markers

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Anti-Hsp90α (ab74248), anti-Hsp90β (ab2927), and anti-Rhodopsin (ab5417) antibodies were from Abcam. Anti-pHsp90α (T5/7) (#3488) antibody was from Cell Signaling Technology. Anti-acetylated α-tubulin (sc-23950) and anti-Ub (sc8017) antibodies were from Santa Cruz. Anti-Rab8 (610844) and anti-GM130 (610822) antibodies were from BD Pharmingen. Anti-Hsp70 (ADI-SPA-810-F) antibody was from Enzo. Anti-γ-tubulin (T6557), anti-α-tubulin (T5168), anti-GAPDH (G8795), anti-Tulp1 (SAB2700274), anti-MAP1B (HPA022275) antibodies, horse radish peroxidase-conjugated secondary antibody, and DAPI were from Sigma. Alexa-Fluor 488- or 546-conjugated secondary antibodies and anti-fade mounting medium were from Molecular Probes/Invitrogen. TUNEL FITC Apoptosis Detection Kit was from Vazyme. 17-DMAG was from Selleck. Protease and phosphatase inhibitors, Protein A/G magnetic beads, and Anti-Flag affinity gel were from Biomake.
Wu Y., Zheng X., Ding Y., Zhou M., Wei Z., Liu T, & Liao K. (2019). The molecular chaperone Hsp90α deficiency causes retinal degeneration by disrupting Golgi organization and vesicle transportation in photoreceptors. Journal of Molecular Cell Biology, 12(3), 216-229.
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2

Protein Extraction and Western Blot Analysis

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Cells were lysed in RIPA buffer (50mM Tris HCl, pH 7.7; 0.15M NaCl; 1mM EDTA; 1% Triton X-100) supplemented with protease inhibitors (Roche) and phosphatases inhibitors (Sigma Aldrich). Protein concentration was measured using Bio-Rad protein assay reagent (Biorad Laboratories, CA). Samples were denatured at 95°C for 10 min in loading buffer then resolved in SDS-PAGE and transferred onto nitrocellulose membranes. Membranes were then blocked in 5% non-fat milk in TBS buffer, 0.1% Tween and then immunoblotted using the following primary antibodies in the specified concentrations: anti-Nedd4l (13690-1-AP, rabbit, Proteintech, 1:1000), anti-Actin (mouse, IGBMC, 1:1000), anti-Akt-pS473 (#4060, rabbit, Cell Signalling, 1:1000), rabbit anti-Akt-pT308 (#2965, rabbit, Cell Signalling, 1:1000) anti-Akt (pan) (#4691, rabbit, Cell Signalling, 1:1000), p-S6S236/236 (#2211, rabbit, Cell Signalling, 1/1000), anti-S6 (#2217, rabbit, Cell Signalling, 1:1000), anti-Ub (sc-8017, mouse, Santa Cruz, 1:250), anti-V5 (R96025, mouse, Invitrogen, 1:5000). All WB experiments consist of at least 3 independent replicates.
Broix L., Jagline H., Ivanova E., Schmucker S., Drouot N., Clayton-Smith J., Pagnamenta A.T., Metcalfe K.A., Isidor B., Louvier U.W., Poduri A., Taylor J.C., Tilly P., Poirier K., Saillour Y., Lebrun N., Stemmelen T., Rudolf G., Muraca G., Saintpierre B., Elmorjani A., Moïse M., Weirauch N.B., Guerrini R., Boland A., Olaso R., Masson C., Tripathy R., Keays D., Beldjord C., Nguyen L., Godin J., Kini U., Nischké P., Deleuze J.F., Bahi-Buisson N., Sumara I., Hinckelmann M.V, & Chelly J. (2016). Mutations in the HECT domain of NEDD4L lead to AKT/mTOR pathway deregulation and cause periventricular nodular heterotopia. Nature genetics, 48(11), 1349-1358.
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3

Western Blot Antibody Panel for Cellular Characterization

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Primary antibodies used were as follows: Anti-Flag (#14793), anti-HA-Tag (#3724), anti-GAPDH (#5174), anti-P53 (#32532), anti-Nanog (#8822), anti-SOX2 (#3579), anti-Caspase 3 (#9662), anti-CD133 (#64326), anti-CD44 (#3570) and anti-EGLN1 (#4835) for WB were purchased from Cell Signaling Technology (USA). anti-EGLN1 (#NB100-2219) for IHC was obtained from Novus Biologicals (USA). Anti-Bax (#50599‐2‐Ig) and anti-Bcl-2 (#12789‐1‐AP) was obtained from Protein Technology (Wuhan, China). Anti-Ub (sc-8017) were purchased from Santa Cruz Biotechnology (CA, USA). Anti-Flag (F1804) were purchased from Sigma Aldrich (USA). Dimethyloxalylglycine (DMOG), carbobenzoxy-Leu-Leu-leucinal (MG132), and cycloheximide (CHX) were purchased from Medchemexpress (USA).
Sun L., Wu C., Ming J., Guo E., Zhang W., Li L, & Hu G. (2022). EGLN1 induces tumorigenesis and radioresistance in nasopharyngeal carcinoma by promoting ubiquitination of p53 in a hydroxylase-dependent manner. Journal of Cancer, 13(7), 2061-2073.
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4

Antibody Validation for Western Blotting and IHC

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The antibodies used in this study were as follows: anti-USP28 (17707-1-AP, 1:1000 WB, 1:200 IHC, Proteintech); anti-MYCN (51705S, 1:1000 WB, 1:200 IHC, Cell Signaling Technology); anti-MYCN (sc-53993, 1:200 WB, Santa Cruz Biotechnology); anti-c-MYC (sc-40, 1:200 WB, Santa Cruz Biotechnology); anti-CyclinD1 (2922S, 1:1000 WB, Cell Signaling); anti-PARP (9542P, 1:1000 WB, Cell Signaling); anti-cleaved PARP (9664S, 1:1000 WB, Cell Signaling); anti-Flag (F3165 1:5000 WB, Sigma); anti-Flag (20543-1-AP, 1:2000 WB, Proteintech); anti-HA (901,503, 1:1000 WB, BioLegend); anti-Ub (SC-8017, 1:200 WB, Santa Cruz Biotechnology); anti-LSD1 (20813-1-AP, 1:1000 WB, Proteintech); anti-Ki67 (27309-1-AP, 1:1000 WB, Proteintech); anti-Actin (66009-1-Ig, 1:5000 WB, Proteintech); and anti-GAPDH (60004-1-Ig, 1:5000 WB, Proteintech); the secondary antibodies conjugated to horseradish peroxidase were used for Western blotting. The secondary antibodies of anti-mouse or anti-rabbit containing Alexa Fluor 488 or 594 were used for immunofluorescence staining (Jackson ImmunoResearch Laboratories).
Li J., Peng J., Wu L., Shen X., Zhen X., Zhang Y., Ma H., Xu Y., Xiong Q., Zhu Q, & Zhang P. (2023). The deubiquitinase USP28 maintains the expression of the transcription factor MYCN and is essential in neuroblastoma cells. The Journal of Biological Chemistry, 299(7), 104856.
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5

Protein Extraction and Western Blot Analysis

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Cells were lysed in RIPA buffer (50mM Tris HCl, pH 7.7; 0.15M NaCl; 1mM EDTA; 1% Triton X-100) supplemented with protease inhibitors (Roche) and phosphatases inhibitors (Sigma Aldrich). Protein concentration was measured using Bio-Rad protein assay reagent (Biorad Laboratories, CA). Samples were denatured at 95°C for 10 min in loading buffer then resolved in SDS-PAGE and transferred onto nitrocellulose membranes. Membranes were then blocked in 5% non-fat milk in TBS buffer, 0.1% Tween and then immunoblotted using the following primary antibodies in the specified concentrations: anti-Nedd4l (13690-1-AP, rabbit, Proteintech, 1:1000), anti-Actin (mouse, IGBMC, 1:1000), anti-Akt-pS473 (#4060, rabbit, Cell Signalling, 1:1000), rabbit anti-Akt-pT308 (#2965, rabbit, Cell Signalling, 1:1000) anti-Akt (pan) (#4691, rabbit, Cell Signalling, 1:1000), p-S6S236/236 (#2211, rabbit, Cell Signalling, 1/1000), anti-S6 (#2217, rabbit, Cell Signalling, 1:1000), anti-Ub (sc-8017, mouse, Santa Cruz, 1:250), anti-V5 (R96025, mouse, Invitrogen, 1:5000). All WB experiments consist of at least 3 independent replicates.
Broix L., Jagline H., Ivanova E., Schmucker S., Drouot N., Clayton-Smith J., Pagnamenta A.T., Metcalfe K.A., Isidor B., Louvier U.W., Poduri A., Taylor J.C., Tilly P., Poirier K., Saillour Y., Lebrun N., Stemmelen T., Rudolf G., Muraca G., Saintpierre B., Elmorjani A., Moïse M., Weirauch N.B., Guerrini R., Boland A., Olaso R., Masson C., Tripathy R., Keays D., Beldjord C., Nguyen L., Godin J., Kini U., Nischké P., Deleuze J.F., Bahi-Buisson N., Sumara I., Hinckelmann M.V, & Chelly J. (2016). Mutations in the HECT domain of NEDD4L lead to AKT/mTOR pathway deregulation and cause periventricular nodular heterotopia. Nature genetics, 48(11), 1349-1358.
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6

Immunoprecipitation of p53 and Ubiquitin

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Cell lysates were collected by 2% SDS lysis buffer and aliquots were used as input. Non-SDS lysis buffer was used to dilute to the rest of lysates. A total of 2 μg of anti-p53 (sc-126, Santa Cruz Biotechnology), anti-Ub (sc-8017, Santa Cruz Biotechnology) or normal IgG (sc-2025, Santa Cruz Biotechnology) was incubated with lysates. Dynabeads (ThermoFisher Scientific) were used for immunoprecipitation according to the manufacturer’s protocol. Immunoblotting was performed as above and anti-mouse IgG VeriBlot for IP secondary antibody (HRP) (ab131368, Abcam, Cambridge, UK) was used.
Wu C.E., Esfandiari A., Ho Y.H., Wang N., Mahdi A.K., Aptullahoglu E., Lovat P, & Lunec J. (2017). Targeting negative regulation of p53 by MDM2 and WIP1 as a therapeutic strategy in cutaneous melanoma. British Journal of Cancer, 118(4), 495-508.
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7

Comprehensive Antibody Assay Protocol

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Assays were performed as described previously62 (link). S6K, pS6K, CREB, tubulin, HA and Flag antibodies were as previously described62 (link). Other antibodies were purchased as follows: anti-LC3 (PM036), MBL International; anti-P62 (610833), BD Biosciences; anti-AMPKα (2532S), anti-pAMPKα (2535S), anti-ULK1 (8054S), anti-pULK1 (Ser555) (5869S), anti-ERK (4695S), anti-pERK (4370S), anti-PLCγ1 (5690P), anti-pPLCγ1 (Tyr783) (14008S), Cell Signaling Technology; anti-TFEB (A303-673A), anti-InsP3R1 (A302-158A), Bethyl Laboratories; anti-PPP1CA (AYA361), anti-PPP1CB (AYA1025), anti-PPP1CC (AYA1169), ayaBIO; anti-calcineurin (A1063), ABclonal Technology; anti-FGFR1 (GTX107393), anti-β-Klotho (GTX45558), anti-InsP3R2 (GTX54772), GeneTex; anti-ASB3 (AP16752A), Abgent; anti-LAMP1 (L1418), anti-MID1 (M2198), Sigma-Aldrich; anti-UBR5 (ab134089), Abcam; anti-GFP (MMS-118P), Covance; anti-Ub (SC-8017), anti-PPP2CA (SC-80665), anti-DREAM (SC-9142), Santa Cruz.
Chen L., Wang K., Long A., Jia L., Zhang Y., Deng H., Li Y., Han J, & Wang Y. (2017). Fasting-induced hormonal regulation of lysosomal function. Cell Research, 27(6), 748-763.
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8

Antibody Validation and Autophagy Modulation

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Anti-Flag (F3165) and anti-HA (H6908) antibodies were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-Myc (sc40), anti-GAPDH (sc47724), anti-BECN1 (sc48341), and anti-Ub (sc-8017) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-TRAF6 (8028S) and anti-LC3 (2775S) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-Mouse IgG H&L (HRP) (ab6728) was purchased from Abcam (Cambridge, MA, USA). Goat anti-Rabbit IgG antibody (HRP) (GTX213110-01) was purchased from GeneTex Inc. (Irvine, CA, USA). Lipopolysaccharide (LPS; L3024), dimethyl sulfoxide (DMSO; 472301), paraformaldehyde (P6148), Triton X-100 (T8787), 3-methyladenine (3-MA; M9281), and Dulbecco’s phosphate-buffered saline (DPBS; D8537) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Polyinosinic-polycytidylic acid (Poly(I:C), tlrl-pic) was purchased from InvivoGen (San Diego, CA, USA). Anti-β-arrestin 2 antibody (PA1-732) and Lipofectamine 2000 (11668019) were purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Kim J.Y., Shin J.H., Kim M.J., Kang Y., Lee J.S., Son J., Jeong S.K., Kim D., Kim D.H., Chun E, & Lee K.Y. (2023). β‐arrestin 2 negatively regulates lung cancer progression by inhibiting the TRAF6 signaling axis for NF-κB activation and autophagy induced by TLR3 and TLR4. Cell Death & Disease, 14(7), 422.
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9

Characterization of CtIP Protein Modifications

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GFP-CtIP, full-length and truncated FLAG–CtIP were generously provided by Dr. Junjie Chen (MD Anderson Cancer Center, TX). FLAG–ASF1A was generously provided by Dr. Zhiguo Zhang (Columbia University, NY). Full-length and truncated FLAG–USP52 and Myc-USP52 were generously provided by Dr. Lei Shi (Tianjin Medical University, Tianjin, China). FLAG–USP52 S1003A and S469A, FLAG–CtIP K62R, K760R, K782R, and 2KR mutants were generated by site-directed mutagenesis (Stratagene). Flag–CtIP–T847E was purchased from Addgene. CHX, IgG agarose, anti-FLAG agarose, 3×FLAG peptide and ATM inhibitor KU55933 were purchased from Sigma Aldrich. Olaparib was purchased from Toronto Research Chemicals. Anti-USP52 (ab241505, 1:1000) was purchased from abcam; anti-CtIP (Thr847) (p1012-847, 1:2000), and anti-CtIP (Thr327) (p1012-327, 1:2000) were purchased from PhosphoSolutions; anti-RPA32 (sc-56770, 1:2000), anti-Ub (sc-8017, 1:2000) and anti-CtIP (sc-271339, 1:1000 for WB) were purchased from Santa Cruz; anti-FLAG (F1804, 1:2000) was purchased from Sigma; anti-pS345 Chk1 (2348, 1:1000), anti-Myc Tag (2276, 1:2000) and anti-SQ/TQ motif (9607, 1:1000) were purchased from CST; anti-CtIP (61141, 1:1000 for IF) was purchased from Active Motif; anti-GAPDH (60004-1-lg, 1:2000) was purchased from Proteintech; anti-Rad51 (GTX100469, 1:1000) was purchased from Genetex.
Gao M., Guo G., Huang J., Kloeber J.A., Zhao F., Deng M., Tu X., Kim W., Zhou Q., Zhang C., Yin P., Luo K, & Lou Z. (2020). USP52 regulates DNA end resection and chemosensitivity through removing inhibitory ubiquitination from CtIP. Nature Communications, 11, 5362.
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10

Immune Signaling Pathway Activation

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LPS (E. coli O111:B4) and poly(I:C) were purchased from Sigma-Aldrich. Pam3CSK4 was purchased from InvivoGen. MG132 was obtained from Merck-Calbiochem. Escherichia coli O111:B4 was obtained from the China Center for Type Culture Collection. Anti-p-IκBα (9246), anti-p-p38 (9215), anti-p-JNK (9251), anti-p-ERK (4377), and anti-p-IRF3 (Ser 396, 29047) were purchased from Cell Signaling Technology. Anti-p38 (A11340), anti-JNK1 (A0288), anti-TLR4 (A5258), anti-MyD88 (A0980), anti-TRIF (A1155), and anti-ERK (A10613) antibodies were purchased from Abclonal Technology. Anti-S100A10 (11250-1-AP), anti-IRF3 (11312-1-AP), and anti-β-actin (66009-1-Ig) were purchased from Proteintech. Anti-Ub (sc-8017) and protein A/G agarose (sc-2003) used for immunoprecipitation were purchased from Santa Cruz Biotechnology. Anti-Flag (F3165) was purchased from Sigma. Anti-HA (901513, previously Covance catalog# MMS-101R) and anti-V5 (680601) were obtained from BioLegend. Fluorochrome-conjugated anti-CD4 (GK1.5), anti-CD8 (53-6.7), anti-B220 (RA3-6B2), anti-Ly-6G (1A8), anti-CD11b (M1/70), anti-CD11c (N418), anti-TLR4 (SA15-21), anti-F4/80 (BM8), anti-CD44 (IM7), and anti-CD62L (MEL-14) antibodies were purchased from BioLegend. Fluorochrome-conjugated anti-CD25 (PC61.5) and Foxp3 (FJK-16s) antibodies were purchased from eBioscience.
Lou Y., Han M., Liu H., Niu Y., Liang Y., Guo J., Zhang W, & Wang H. (2019). Essential roles of S100A10 in Toll-like receptor signaling and immunity to infection. Cellular and Molecular Immunology, 17(10), 1053-1062.
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