C myc y69

Manufactured by Abcam

Sourced in United States

C-Myc (Y69) is a monoclonal antibody that recognizes the c-Myc protein. The c-Myc protein is a transcription factor involved in the regulation of cell growth and proliferation.

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Lab products found in correlation

β actin, by Santa Cruz Biotechnology (2 mentions) N myc ncm 2 100, by Abcam (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Cd138 281 2, by BD (1 mentions) Hemavet hv950fs, by Drew Scientific (1 mentions) Gfap g a 5, by Merck Group (1 mentions) Olig2, by Merck Group (1 mentions) Lamin b1, by Abcam (1 mentions) β tubulin, by Cell Signaling Technology (1 mentions) Hsp70, by Abcam (1 mentions) Cyclophilin b, by Cell Signaling Technology (1 mentions) Ki67 sp6, by Abcam (1 mentions) Cdkn2a p16ink4a, by Abcam (1 mentions) P histone h3, by Cell Signaling Technology (1 mentions) Bcl2 124, by Cell Signaling Technology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Phospho p38 mapk thr180 tyr182, by Cell Signaling Technology (1 mentions) Ephb4, by Abcam (1 mentions) Phospho eif2α s51, by Cell Signaling Technology (1 mentions) Glut3, by Abcam (1 mentions)

Spelling variants of this product

C-Myc (362 citations) MYC (Y69) (8 citations) Anti-c-MYC-Y69 (8 citations) C-MYC (clone Y69, (4 citations)

8 protocols using c myc y69

Peripheral Blood and Tissue Analysis

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Peripheral blood was collected from tail veins and analyzed on an automated blood
counter, HEMAVET HV950FS (Drew Scientific). Tissue samples were fixed
immediately after isolation and processed into paraffin, sectioned and examined
histologically using hematoxylin and eosin, Congo red, or immunohistochemical
techniques. Immunohistochemical staining was performed using the following
anti-mouse antibodies: CD138 (281-2, BD Pharmingen), λ
(SouthernBiotech), κ (SouthernBiotech), and c-Myc (Y-69, Abcam).
Samples were reviewed by pathologists and diagnosed using uniform criteria.

Asai T., Hatlen M.A., Lossos C., Ndiaye-Lobry D., Deblasio A., Murata K., Fleisher M., Cortizas E.M., Verdun R.E., Petrini J, & Nimer S.D. (2016). Generation of a novel, multi-stage, progressive, and transplantable model of plasma cell neoplasms. Scientific Reports, 6, 22760.

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Antibody Characterization for Immunostaining

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Antibodies for immunostaining and western blotting were used as per the manufacturer’s recommendations. The following antibodies were used within this study: β-tubulin (Cell Signalling Technology, Cat # 2146 (1:1000)), Cleaved Caspase-3 [Asp175] (Abcam, Cat # ab49899 (WB 1:500, IHC 1:1500)), CDKN2A/p19ARF (Abcam, Cat # ab80 (1:1000)), cMYC [Y69] (Abcam, Cat # ab32072 (WB 1:1000, IHC 1:500)), Cyclophilin B (Cell Signalling Technology, Cat # 43603 (1:1000)), GFAP [GA5] (Millipore, Cat # MAB3402 (1:1000)), GFP (Abcam, Cat # ab13970 (1:1000)), HSF1 (Abcam, Cat # ab61382 (1:1000)), HSP70 (Abcam, Cat # ab2787 (1:1000)), HSP90 [EPR16621] (Abcam, Cat # ab203085 (1:1000)), Ki67 [SP6] (Abcam, Cat # ab16667 (1:2000)), n-MYC [NCM II 100] (Abcam, Cat # ab16898 (1:250)), NPR3 (Abcam, Cat # ab97389 (1:250)), Olig-2 (Millipore, Cat # AB9610 (1:1000)), OTX2 (R&D Systems, Cat # AF1979 (1:500)), p21 (Abcam, Cat # ab109199 (1:1000)), Synaptophysin [SY38] (Millipore, Cat # MAB5258 (1:500)), TUJ1 (Covance, Cat # MMS-435P (1:500)), β-actin (Santa Cruz Biotechnology, Cat # sc-47778 (1:1000)), CDKN2A/p16INK4A (Abcam, Cat # ab211542 (1:1000)), anti-mouse IgG secondary HRP (VWR, Cat # NXA931 (1:1000)), anti-rabbit IgG secondary HRP (GE Healthcare, Cat # NA934 (1:1000)), and Lamin B1 (Abcam, Cat # ab16048 (1:1000)).

Mainwaring O.J., Weishaupt H., Zhao M., Rosén G., Borgenvik A., Breinschmid L., Verbaan A.D., Richardson S., Thompson D., Clifford S.C., Hill R.M., Annusver K., Sundström A., Holmberg K.O., Kasper M., Hutter S, & Swartling F.J. (2023). ARF suppression by MYC but not MYCN confers increased malignancy of aggressive pediatric brain tumors. Nature Communications, 14, 1221.

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Phospho-Aurora Kinase Signaling Assay

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Cells were treated with 100nM MLN8237 and 1mM cyclophosphamide for 24-48h. To detect Phospho-Aurora Kinases, cells were treated with 60nM nocodazole for 16h, followed by 100nM MLN8237 and/or 1mM cyclophosphamide for 2h. Lysate was obtained, and equivalent amounts of protein were loaded onto 10% SDS-PAGE gels. The gels were transferred to nitrocellulose and incubated with primary and secondary antibodies purchased from Cell Signaling Technology, as previously described. Primary antibodies used: Mdm2 (D12), p53 (DO-7), actin (2Q1055; Santa Cruz Biotechnology, Inc.; Dallas, TX, USA), c-Myc (Y69; Abcam, Cambridge, UK), P-Rb (D20B12), Mdr1 (E1Y7B), AurkA (D3V7T), AurkB (3094), P-AurkA/B/C (D13A11), P-Histone H3 (Ser10; D7N8E), P-Src Family Y416 (D49G4), LC3B (E5Q2K), Bcl2 (124), Bax (D2E11), GAPDH (D16H11), AIF (D39D2), TBP (D5C9H), Caspase 3 (9662), Caspase 7 (C7), tubulin (2148; Cell Signaling Technology, Danvers, MA, USA).

Park S.I., Lin C.P., Ren N., Angus S.P., Dittmer D.P., Foote M., Parton T., Bhatt A.P., Fedoriw Y.D., Roth D.P., Cann M.L., Johnson G.L, & Damania B. (2019). Inhibition of Aurora A Kinase in Combination with Chemotherapy Induces Synthetic Lethality and Overcomes Chemoresistance in Myc-overexpressing Lymphoma. Targeted oncology, 14(5), 563-575.

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Western Blot Analysis of Signaling Proteins

Cited 2 times

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This was performed as described^{56 (link),57 (link)}. The following primary antibodies were used: EPHB4 (D1C7N) (1: 1000, Cell Signaling #14960), EPHB4 (1:500, Abcam #ab73259), phospho-eiF2α S51 (1:1000, Cell Signaling #9721 S), eiF2α (1:000, Cell Signaling #9722), phospho-IRE1(S724) (1:1000, Abcam #ab48187), c-Myc (Y69) (1:5000, Abcam #ab32072), GLUT3 (1:5000, Abcam #ab191071), Calreticulin (1:400, Abcam #ab2907), HMGB1 (Abcam #ab18256), Actin (1:10000, Cell Signaling #5125), GAPDH (1:5000, Cell Signaling #3683), Phospho-Src Family Tyr416 (1:1000, Cell Signaling #2101), Phospho-p38 MAPK Thr180/Tyr182 (1:1000, Cell Signaling #9211), and Phospho-4EBP1 Thr37/46 (236B4) (1:1000, Cell Signaling #2855). Following incubation with horseradish peroxidase-conjugated goat anti-rabbit/mouse (1:5000, Biorad) or goat anti-rat (1:5000, Santa Cruz) for 1 h, the binding of secondary antibodies were detected with the Supersignal West Femto Maximum Sensitivity Substrate detection system (Pierce) followed by visualization. Quantification analyses were performed by Biorad ChemiDoc Imager and Bio-Rad Image Lab software.

Sagar V., Vatapalli R., Lysy B., Pamarthy S., Anker J.F., Rodriguez Y., Han H., Unno K., Stadler W.M., Catalona W.J., Hussain M., Gill P.S, & Abdulkadir S.A. (2019). EPHB4 inhibition activates ER stress to promote immunogenic cell death of prostate cancer cells. Cell Death & Disease, 10(11), 801.

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Western Blot Analysis of Protein Signaling

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Cells and xenograft tumor samples were resuspended in high SDS-RIPA Buffer (50 mM Tris-HCl, pH 7.5, 150 mM Sodium Chloride, 1 % Triton X-100, 1 % sodium deoxycholate, 1 % SDS, 2 mM EDTA; Sigma Aldrich). Tissues were disrupted and homogenized with a TissueLyser II (Qiagen) for 2 × 2 min intervals at 30Hz. Protein concentration was determined using the Pierce™ BCA Protein Assay Kit (Pierce). A total of 15–50 μg of protein extracts were loaded onto NuPAGE® Novex® 4–12 % Bis-Tris Protein Gels (Life Technologies) and subsequently transferred onto nitrocellulose membranes using the iBlot® Dry Blotting System (Life Technologies). The blots were developed using SuperSignal™ West Pico Chemiluminescent Substrate (Thermo Scientific). Antibodies: S6-Ribosomal protein (5G10), Phospho-S6 Ribosomal protein (Ser240/244) (D68F8), Phospho-4E-BP1 (Thr37/46) (236B4), p44/42 MAPK (Erk1/2) (137 F5), and Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) were purchased from Cell Signaling Technology. C-MYC (Y69) and N-MYC (NCM II 100) were purchased from Abcam. FLAG (M2) and β-actin (A2066) antibodies were purchased from Sigma Aldrich.

Dela Cruz F.S., Diolaiti D., Turk A.T., Rainey A.R., Ambesi-Impiombato A., Andrews S.J., Mansukhani M.M., Nagy P.L., Alvarez M.J., Califano A., Forouhar F., Modzelewski B., Mitchell C.M., Yamashiro D.J., Marks L.J., Bender J.L, & Kung A.L. (2016). A case study of an integrative genomic and experimental therapeutic approach for rare tumors: identification of vulnerabilities in a pediatric poorly differentiated carcinoma. Genome Medicine, 8, 116.

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Chemotherapeutic Agents and Antibody Procurement

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TA3 (Figure 1A) was purchased from Chemfaces (Wuhan Chemfaces Biochemical Co., Ltd., China). DOX was purchased from Selleckchem (Munich, Germany), while 5-FU was purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies for CNOT2 (Cat No. 34214) and PARP (Cat No. 9542) were purchased from Cell Signaling Technology (Beverly, MA, USA), MID1IP1 (Cat No.15764-1) was purchased from Proteintech (Rosemont, IL, USA), and c-Myc(Y69) (Cat No. 32072) was purchased from Abcam (Cambridge, UK). Caspase 3 (Cat No. sc-7272), β-actin (Cat No. sc-47778), and secondary antibodies (Cat No. sc-516102; sc-2004) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Ko H.M., Jee W., Park D.I., Kim K.I., Jung J.H, & Jang H.J. (2022). The Antitumor Effect of Timosaponin A3 through c-Myc Inhibition in Colorectal Cancer Cells and Combined Treatment Effect with 5-FU or Doxorubicin. International Journal of Molecular Sciences, 23(19), 11900.

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Comprehensive Immune Cell Phenotyping

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The following antibodies were used: APC labeled PBS57 loaded CD1d-tetramers from the NIH tetramer facility, TCRβ APC-e780 (H57-597, eBiosciences) or Pacific Blue (H57-597, Biolegend), CD44 AlexaFluor(AF)700 (IM7, Biolegend), CD45.2 Pacific Blue (104, Biolegend), Thy1.1 phycoerythrin (PE) (A85-1, BD Pharmingen), Thy1.2 FITC (53-2.1, BD Pharmingen), NK1.1 PE-Cy7 or PE (PK136, BD Pharmingen), or Pacific Blue (PK136, Biolegend), or NK1.1-biotin (PK136, eBiosciences) followed by streptavidin-PE Texas Red, CD24 PerCPCy5.5 (M1/69, eBiosciences) or FITC or PE (M1/69, BD Pharmingen), IL-17A PE (17B7, eBiosciences), IL-4 PE-Cy7 (BVD6-24G2, eBiosciences), IFN-γ PerCPCy5.5 (XMG1.2, Biolegend), cMyc (Y69, abcam) or p27^kip1 (D69C12; Cell Signaling Technology) followed by anti-rabbit IgG PE (Life Sciences), Ki67-PeCy7 (B56, BD Pharmingen), rabbit IgG isotype (Life Technologies) and BrdU (BrdU flow kit; BD Pharmingen). For transcription factor staining, cells were incubated with antibody to PLZF (D-9, Santa Cruz), followed by anti-mouse IgG1 PE (A85-1, eBiosciences), and then T-bet PE-Cy7 (4B10, eBiosciences), GATA3 PerCpCy5.5 (TWAJ, eBiosciences), and RORγt-BV421 (Q31-378, BD Biosciences).

Sklarz T., Guan P., Gohil M., Cotton R.M., Ge M.Q., Haczku A., Das R, & Jordan M.S. (2017). mTORC2 regulates multiple aspects of NKT-cell development and function. European journal of immunology, 47(3), 516-526.

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Western Blot Analysis of Uhrf1, c-Myc, and AP4

Cited 1 time

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Western blot analysis was performed as previously described (Muppidi et al., 2014 (link)). In brief, cell lysates were separated by SDS-PAGE. Proteins were detected with antibodies against Uhrf1 (M-132; Santa Cruz Biotechnology), c-Myc (Y69; Abcam), AP4 (A-8; Santa Cruz Biotechnology), and β-actin (Cell Signaling Technology).

Chen C., Zhai S., Zhang L., Chen J., Long X., Qin J., Li J., Huo R, & Wang X. (2018). Uhrf1 regulates germinal center B cell expansion and affinity maturation to control viral infection. The Journal of Experimental Medicine, 215(5), 1437-1448.

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