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C myc clone 9e10

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C-Myc (clone 9E10) is a mouse monoclonal antibody that recognizes the c-Myc protein. The c-Myc protein is a transcription factor that plays a crucial role in cell growth, proliferation, and apoptosis. The C-Myc (clone 9E10) antibody is commonly used in various research applications, including immunoprecipitation, immunoblotting, and immunohistochemistry.

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Lab products found in correlation

Glut1, by Abcam (3 mentions) Tubulin, by Thermo Fisher Scientific (2 mentions) Alkbh5, by Abcam (2 mentions) Mettl3, by Cell Signaling Technology (2 mentions) O glcnac clone rl2, by Thermo Fisher Scientific (2 mentions) D2hgdh, by Proteintech (2 mentions) L2hgdh, by Proteintech (2 mentions) Fto c 3, by Santa Cruz Biotechnology (2 mentions) Mettl14, by Cell Signaling Technology (2 mentions) Lamin a, by Fortis Life Sciences (2 mentions) β actin, by Merck Group (2 mentions) Anti human melan a clone a103, by Agilent Technologies (2 mentions) Rabbit igg, by Vector Laboratories (2 mentions) P44 42 mapk erk1 2, by Cell Signaling Technology (2 mentions) Cyclind1 clone ep12, by Agilent Technologies (2 mentions) Mouse igg, by Vector Laboratories (2 mentions) Phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions) β catenin, by BD (2 mentions) 3 3 diaminobenzidine (dab), by Agilent Technologies (2 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)

11 protocols using c myc clone 9e10

1

Immunohistochemical Analysis of Tumor Markers

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Immunohistochemistry was performed as described previously [35 (link)]. Tumor sections prepared from control and silibinin-treated groups were stained with Ki67 (Thermo Fisher Scientific, Waltham, MA, USA) and c-Myc (clone 9E10; Santa Cruz Biotechnology, Dallas, Texas, USA), pSTAT3 (Cell Signaling Technology, Danvers, MA, USA) and GLUT1 (Abcam, Cambridge, UK) primary antibodies. Images were captured at 20X magnification using an inverted microscope (Leica, DMI600B) and processed by using Leica LAS AF software.
Shukla S.K., Dasgupta A., Mehla K., Gunda V., Vernucci E., Souchek J., Goode G., King R., Mishra A., Rai I., Nagarajan S., Chaika N.V., Yu F, & Singh P.K. (2015). Silibinin-mediated metabolic reprogramming attenuates pancreatic cancer-induced cachexia and tumor growth. Oncotarget, 6(38), 41146-41161.
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2

Comprehensive Protein Analysis in Metabolic Disorders

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Whole-cell lysates (WCL) were examined as we described36 (link). Nuclear and cytoplasmic proteins were extracted and analyzed as we recently reported5 (link). The following antibodies were used: c-MYC (clone 9E10; #sc-40, Santa Cruz Biotechnology), IDH2 (#ab55271, Abcam), OGT (#24083, Cell Signaling Technology), OGA (Cat #14711–1-AP, Proteintech), O-GlcNAc (clone RL2, #MA1–072, Thermo-Fisher Scientific), D2HGDH (#13895–1-AP, Proteintech), L2HGDH (#15707–1-AP, Proteintech), ALKBH5 (# ab69325, Abcam), FTO (c-3, #sc-271713, Santa Cruz Biotechnology), TET1 (#SAB2700730, Sigma-Aldrich), TET2 (#18950, Cell Signaling Technology), TET3 (#: ABS463, EMD Millipore Corporation), METTL3, METTL14, WTAP (# 69391, # 51104 and #56501, all from Cell Signaling Technology), β-actin (# A2228, Sigma-Aldrich), Lamin A (#A303–433A-M, Bethyl Laboratories), Tubulin (#62204 Invitrogen). PVDF membranes were stripped and re-probed with relevant antibodies for loading control, as reported37 (link).
Lin A.P., Qiu Z., Ethiraj P., Sasi B., Jaafar C., Rakheja D, & Aguiar R.C. (2022). MYC, mitochondrial metabolism and O-GlcNAcylation converge to modulate the activity and subcellular localization of DNA and RNA demethylases. Leukemia, 36(4), 1150-1159.
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3

Comprehensive Protein Analysis in Metabolic Disorders

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Whole-cell lysates (WCL) were examined as we described36 (link). Nuclear and cytoplasmic proteins were extracted and analyzed as we recently reported5 (link). The following antibodies were used: c-MYC (clone 9E10; #sc-40, Santa Cruz Biotechnology), IDH2 (#ab55271, Abcam), OGT (#24083, Cell Signaling Technology), OGA (Cat #14711–1-AP, Proteintech), O-GlcNAc (clone RL2, #MA1–072, Thermo-Fisher Scientific), D2HGDH (#13895–1-AP, Proteintech), L2HGDH (#15707–1-AP, Proteintech), ALKBH5 (# ab69325, Abcam), FTO (c-3, #sc-271713, Santa Cruz Biotechnology), TET1 (#SAB2700730, Sigma-Aldrich), TET2 (#18950, Cell Signaling Technology), TET3 (#: ABS463, EMD Millipore Corporation), METTL3, METTL14, WTAP (# 69391, # 51104 and #56501, all from Cell Signaling Technology), β-actin (# A2228, Sigma-Aldrich), Lamin A (#A303–433A-M, Bethyl Laboratories), Tubulin (#62204 Invitrogen). PVDF membranes were stripped and re-probed with relevant antibodies for loading control, as reported37 (link).
Lin A.P., Qiu Z., Ethiraj P., Sasi B., Jaafar C., Rakheja D, & Aguiar R.C. (2022). MYC, mitochondrial metabolism and O-GlcNAcylation converge to modulate the activity and subcellular localization of DNA and RNA demethylases. Leukemia, 36(4), 1150-1159.
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4

Immunohistochemical Analysis of Melanoma Markers

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Briefly, formalin fixed, paraffin embedded sections were deparaffinised, rehydrated, blocked for endogenous peroxidases and underwent antigen retrieval according to antibody specifications. Tissues were incubated overnight with the following primary antibodies. Anti-human melan-a clone A103 (Dako, M7196), S100 (Dako, Z0311), β-catenin (BD Transduction Laboratories, 610154), CyclinD1 clone EP12 (Dako, M3642), c-myc clone 9E10 (Santa Cruz Biotechnology, sc-40), p44/42 MAPK (Erk1/2) (Cell Signaling, 4695) and phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling, 9101), mouse IgG (Vector, I-2000), and rabbit IgG (Vector, I-1000). Secondary antibodies used for DAB based IHC were either EnVision+ System- HRP Labelled Polymer Anti-mouse (Dako, K4001) or EnVision+ System- HRP Labelled Polymer Anti-rabbit (Dako, K4003) based on primary antibody host species. Peroxidase activity was revealed using DAB (Dako, K3468). Samples were then counterstained with haematoxylin, dehydrated, and coverslipped.
Pawlikowski J.S., Brock C., Chen S.C., Al-Olabi L., Nixon C., McGregor F., Paine S., Chanudet E., Lambie W., Holmes W.M., Mullin J.M., Richmond A., Wu H., Blyth K., King A., Kinsler V.A, & Adams P.D. (2015). Acute Inhibition of MEK Suppresses Congenital Melanocytic Nevus Syndrome in a Murine Model Driven by Activated NRAS and Wnt Signaling. The Journal of Investigative Dermatology, 135(8), 2093-2101.
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5

Protein Isolation and Western Blotting

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Protein isolation and western blotting were performed as described previously [49 (link)]. Briefly, cells were washed twice with PBS and lysed in radioimmuno precipitation assay (RIPA) lysis buffer by incubating at 4°C on a rotatory shaker for 30 min. To remove the cell debris, lysates were centrifuged at 13000 rpm for 10 min. Protein concentration was measured by Bradford assay. Equal amount of protein was used for western blotting. Primary antibodies against GLUT1 (Abcam, Cambridge, UK), c-Myc (clone 9E10; Santa Cruz Biotechnology, Dallas, Texas, USA), HKII, LDHA, STAT3, pSTAT3, anti-phospho histone H2A. X (Cell Signaling Technology, Danvers, MA, USA) and actin (Developmental Studies Hybridoma Bank, Iowa City, IA) were utilized for probing specific proteins.
Shukla S.K., Dasgupta A., Mehla K., Gunda V., Vernucci E., Souchek J., Goode G., King R., Mishra A., Rai I., Nagarajan S., Chaika N.V., Yu F, & Singh P.K. (2015). Silibinin-mediated metabolic reprogramming attenuates pancreatic cancer-induced cachexia and tumor growth. Oncotarget, 6(38), 41146-41161.
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6

Western Blotting Analysis of Protein Expression

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Western blotting of proteins was performed as described previously [20 (link)]. Briefly, for western blotting, cells were washed twice with cold PBS and lysed in radioimmuno precipitation assay (RIPA) lysis buffer by incubating at 4°C on a rotatory shaker for 30 min. Cell debris was removed by centrifugation at 13,000 rpm for 10 min and the supernatant was collected. Protein content was measured by Bradford assay. Equal amounts of denatured proteins were separated by electrophoresis using SDS-PAGE gels and transferred to activated PVDF membranes. The membranes were probed with primary antibodies against GLUT1 (Abcam, Cambridge, UK), c-Myc (clone 9E10; Santa Cruz Biotechnology, Dallas, Texas, USA), HKII, pP70S6K, P70S6K, p4EBP1, 4EBP1 (Cell Signaling Technology, Danvers, MA, USA) and beta-tubulin (Clone E7 from Developmental Studies Hybridoma Bank, Iowa City, IA).
Shukla S.K., Gunda V., Abrego J., Haridas D., Mishra A., Souchek J., Chaika N.V., Yu F., Sasson A.R., Lazenby A.J., Batra S.K, & Singh P.K. (2015). MUC16-mediated activation of mTOR and c-MYC reprograms pancreatic cancer metabolism. Oncotarget, 6(22), 19118-19131.
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7

Western Blotting of Cell Signaling Proteins

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For western blotting, cells were washed and harvested in ice-cold PBS and lysed in radioimmunoprecipitation assay (RIPA) buffer containing 10 mM protease inhibitor cocktail (Thermo Scientific, Lafayette, CO, USA). The cell lysates were then separated on NuPAGE 4%–12% gradient gels (Invitrogen, Carlsbad, CA, USA), proteins were transferred to nitrocellulose membrane (Bio-Rad, Hercules, CA, USA) and incubated with primary antibodies overnight, followed by incubation with horseradish-peroxidase (HRP)-conjugated secondary antibodies and visualization with enhanced chemiluminescence reagent (Thermo Scientific, Lafayette, CO, USA). Antibodies from Cell Signaling (Danvers, MA, USA) were PIM-3 (clone D17C9), p-Bad S112 (clone S112), Bcl-xL (clone 54HS), p-Stat3 Y705 (clone D3A7), p-MDM2 S166 (cat. no. 3521S), p21 Waf/Cip1 (clone 12D1), Cdc25a (cat no. 3652S), CDK2 (clone 78B2), cyclin D1 (clone 92G2), cyclin E1 (clone HE12), cyclin A2 (clone BF683), and p-RB Ser795 (cat. no. 9301). Antibodies from Santa Cruz (Dallas, TX, USA) were DHODH (clone E-8), PIM-1 (clone 12H8), PIM-2 (clone 1D12), and c-Myc (clone 9E10). Actin (clone AC-15) was purchased from Millipore-Sigma (Burlington, MA, USA).
Buettner R., Morales C., Wu X., Sanchez J.F., Li H., Melstrom L.G, & Rosen S.T. (2019). Leflunomide Synergizes with Gemcitabine in Growth Inhibition of PC Cells and Impairs c-Myc Signaling through PIM Kinase Targeting. Molecular Therapy Oncolytics, 14, 149-158.
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8

Antibody Procurement and Chemical Reagent Acquisition

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Antibodies were obtained as follows: p-PLK1 (Thr210) (Cell Signalling Technology); Aurora A (Millipore); Aurora B (Abcam); PLK1 (clone F-8), VCAM-1 (clone H-276), ICAM-1 (clone H-108), Bcl-2 (clone N-19), Bax (clone N-20), survivin (clone FL-142), p21 (clone C-19), cyclin B1 (clone GNS1), Wee1 (clone C-20), CDC25 (clone C-20), c-Myc (clone 9E10) and β-actin (Santa Cruz Biotechnology).
Tivozanib was purchased from AdooQ BioScience (Irvine, CA, USA). Gefitinib (EGFR small molecule inhibitor) and temozolomide (a DNA alkylating agent) were obtained from ChemieTek (Indianapolis, IN, USA). Irinotecan (a topoisomerase I inhibitor) was purchased from pharmacy of Shariati hospital (Tehran, Iran). Poly-hydroxyethylmethacrylate polymer (poly-HEMA) was obtained from Santa Cruz Biotechnology.
Momeny M., Moghaddaskho F., Gortany N.K., Yousefi H., Sabourinejad Z., Zarrinrad G., Mirshahvaladi S., Eyvani H., Barghi F., Ahmadinia L., Ghazi-Khansari M., Dehpour A.R., Amanpour S., Tavangar S.M., Dardaei L., Emami A.H., Alimoghaddam K., Ghavamzadeh A, & Ghaffari S.H. (2017). Blockade of vascular endothelial growth factor receptors by tivozanib has potential anti-tumour effects on human glioblastoma cells. Scientific Reports, 7, 44075.
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9

Cell Culture and Viability Assay Protocol

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BSC40 cells (Cat# CRL-2761) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Murine embryonic fibroblasts (MEFs) were a kind gift from Dr. Carl Atkinson. LLC-A9F1 (A9F1) cells were a kind gift from Dr. Mark Rubenstein. All cells were cultured in DMEM supplemented with 10% Fetal Bovine Serum and 1x Penicillin-Streptomycin-L-Glutamine (Corning, Oneonta, NY, USA). Cell viability was measured using the CellTiter-96 Non-Radioactive Cell Proliferation (MTT) assay (Promega, Madison, WI, USA) according to the manufacturer’s recommendations. Antibodies used for this study were purchased from Santa Cruz Biotech (Santa Cruz, CA) and include: actin (clone I19) and c-Myc (clone 9E10).
Burton C., Bartee M.Y, & Bartee E. (2019). Impact of Induced Syncytia Formation on the Oncolytic Potential of Myxoma Virus. Oncolytic Virotherapy, 8, 57-69.
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10

Immunohistochemistry of Formalin-Fixed Paraffin-Embedded Tissues

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Briefly, formalin fixed, paraffin embedded sections were deparaffinised, rehydrated, blocked for endogenous peroxidases, and underwent antigen retrieval according to antibody specifications. Tissues were incubated overnight with the following primary antibodies: Anti-human melan-a clone A103 (Dako, M7196), S100 (Dako, Z0311), β-catenin (610154, BD Transduction Laboratories, San Jose, CA), CyclinD1 clone EP12 (Dako, M3642), c-myc clone 9E10 (Santa Cruz Biotechnology, sc-40), p44/42 MAPK (Erk1/2; Cell Signaling, 4695) and phospho-p44/42 MAPK (Erk1/2, Thr202/Tyr204, Cell Signaling, 9101), mouse IgG (I-2000, Vector, Burlingame, CA), and rabbit IgG (Vector, I-1000). Secondary antibodies used for DAB-based IHC were either EnVision+ System- horseradish peroxidase Labeled Polymer Anti-mouse (Dako, K4001) or EnVision+ System- horseradish peroxidase Labeled Polymer Anti-rabbit (Dako, K4003) based on primary antibody host species. Peroxidase activity was revealed using DAB (Dako, K3468). Samples were then counterstained with haematoxylin, dehydrated, and coverslipped.
Pawlikowski J.S., Brock C., Chen S.C., Al-Olabi L., Nixon C., McGregor F., Paine S., Chanudet E., Lambie W., Holmes W.M., Mullin J.M., Richmond A., Wu H., Blyth K., King A., Kinsler V.A, & Adams P.D. (2015). Acute Inhibition of MEK Suppresses Congenital Melanocytic Nevus Syndrome in a Murine Model Driven by Activated NRAS and Wnt Signaling. The Journal of Investigative Dermatology, 135(8), 2093-2101.
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