Bcl 2 d17c4

Manufactured by Cell Signaling Technology

Sourced in France, China, United States

Bcl-2 (D17C4) is a primary antibody that recognizes the Bcl-2 protein. Bcl-2 is a key regulator of apoptosis (programmed cell death) and plays a crucial role in cell survival and proliferation.

Automatically generated - may contain errors

Lab products found in correlation

Ab110321, by Abcam (1 mentions) Nitrotyrosine, by Merck Group (1 mentions) Ab31163, by Abcam (1 mentions) Ab54481, by Abcam (1 mentions) Ab5605, by Merck Group (1 mentions) Image lab software, by Bio-Rad (1 mentions) Caspase 3 8g10, by Cell Signaling Technology (1 mentions) Bsa protein assay kit, by Thermo Fisher Scientific (1 mentions) Ecl western blotting substrate, by Bio-Rad (1 mentions) Restore western blot stripping buffer, by Thermo Fisher Scientific (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Caspase 9, by Affinity Biosciences (1 mentions) Annexin 5 fitc and propidium iodide kit, by BD (1 mentions) Cdk4 d9g3e, by Cell Signaling Technology (1 mentions) Parp 46d11, by Cell Signaling Technology (1 mentions) Cyclin d1 e3p5s, by Cell Signaling Technology (1 mentions) Bax d2e11, by Cell Signaling Technology (1 mentions) Ros assay kit, by Beyotime (1 mentions) Alexa flour 488, by Abcam (1 mentions)

4 protocols using bcl 2 d17c4

Western Blot Analysis of Protein Targets

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Western Blotting was carried out using specific antibodies; Bcl-2 (D17C4, Cell Signaling, France), Bax (ab115193, Abcam, France), aconitase 2 (ab110321, Abcam, France), nitrotyrosine (05-233, Millipore, France), 4HNE (ab5605, Millipore, France), sirtuine-3 (D22A3, Cell signaling, France), sirtuine-1 (SC-9475, Cell signaling, France), phosphorylated sirtuine-1 (Sc-2314, Cell signaling, France), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1a, ab54481, Abcam, France), nuclear factor (erythroid-derived 2)-like 2 (NRF2, ab31163, Abcam, France), mammalian target of rapamycin (mTOR, 2983S, Cell signaling, France), phospho mTOR (5536P, Cell signaling, France), eNOS total (610296, BD Transduction, Le Pont de Claix, France) and eNOS phophorylated Ser1177 (C9C3; Cell-Signaling, Saint Quentin-Yvelines, France). Protein quantification level was expressed as ratio of total proteins loaded on precast gels with stain-free technology (BioRad, France). All blots were obtained by imaging with chemidoc and quantificated with ImageLab software (BioRad).

Kerforne T., Giraud S., Danion J., Thuillier R., Couturier P., Hebrard W., Mimoz O, & Hauet T. (2019). Rapid or Slow Time to Brain Death? Impact on Kidney Graft Injuries in an Allotransplantation Porcine Model. International Journal of Molecular Sciences, 20(15), 3671.

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Protein Analysis by Western Blotting

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Protein was isolated and concentration quantified using a BSA Protein Assay Kit (ThermoScientific). Ten-20 μg of total protein was separated on a Mini-PROTEAN TGX, Any kDTM gel followed by transfer onto PVDF membranes (Bio-Rad). The membranes were blocked (5% non-fat dry milk in TBS + 0.5% Tween20) prior to overnight incubation at 4 °C with primary antibodies to β-catenin (BD), Bcl-2 (D17C4), Caspase-3 (8G10), or GAPDH (D16HH, Cell Signaling). Following washes, membranes were incubated for 1 h at room temperature with secondary HRP-conjugated goat antibodies to mouse or rabbit IgG (Cell Signaling). Chemiluminescence was performed using Western ECL Blotting Substrate (Bio-Rad) followed by X-ray film-based imaging. Films were scanned and quantified for integrated optical density (IOD) using ImageJ software. To remove antibodies, membranes were incubated for 15 min at room temperature in Restore Western Blot Stripping Buffer (ThermoScientific).

Bailey S.R., Nelson M.H., Majchrzak K., Bowers J.S., Wyatt M.M., Smith A.S., Neal L.R., Shirai K., Carpenito C., June C.H., Zilliox M.J, & Paulos C.M. (2017). Human CD26high T cells elicit tumor immunity against multiple malignancies via enhanced migration and persistence. Nature Communications, 8, 1961.

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Synthesis and Evaluation of Compounds 1-3

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Compounds 1, 2, and 3 were synthesized according to the procedure developed by our group [12 (link)]. They were dissolved in dimethyl sulfoxide (DMSO), the concentration of which never exceeded 0.1% (v/v); 50 mM of stock solution was stored at −20 °C. The Cell counting Kit-8 (CCK-8), Hoechst 33258, JC-1, and the ROS assay kit were purchased from Beyotime Institute of Biotechnology Company (Shanghai, China). The annexin V-FITC and propidium iodide (PI) kit was obtained from BD Pharmingen (BD, San Diego, CA, USA). The primary antibodies used are as follows: CDK4 (D9G3E, Cell Signaling Technology, Danvers, MA, USA), Cyclin D1 (E3P5S, Cell Signaling Technology), Caspase-3 and cleaved Caspase-3 (D3R6Y, Cell Signaling Technology), PARP (46D11, Cell Signaling Technology), Cytochrome c (AF0146, Affinity, Changzhou, China), Caspase-9 (AF6348, Affinity), cleaved Caspase-9 (D8I9E, Cell Signaling Technology), BAX (D2E11, Cell Signaling Technology), Bcl-2 (D17C4, Cell Signaling Technology), Bad (D24A9, Cell Signaling Technology), and β-actin (AF0003, Beyotime Biotechnology, Shanghai, China). All other chemicals used are commercially available and reagent grade.

Zheng J., Zeng L., Tang M., Lin H., Pi C., Xu R, & Cui X. (2021). Novel Ferrocene Derivatives Induce G0/G1 Cell Cycle Arrest and Apoptosis through the Mitochondrial Pathway in Human Hepatocellular Carcinoma. International Journal of Molecular Sciences, 22(6), 3097.

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Immunoblotting and Flow Cytometry Analysis

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All reagents used for media, PBS, and Percoll were obtained from Sigma-Aldrich and BD Bioscience. The primary antibodies
used in these studies included rabbit monoclonal antibodies against phospo-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cell Signaling
#9101, Danvers, MA, USA), phospho-SAPK/JNK (Thre183/Tyr185) (Cell Signaling #4668), phospho-p38 MAPK (Thr180/Tyr182) (Cell
Signaling #9221), phospho-NF-κB p65 (Ser436) (93H1) (Cell Signaling #3033), Cleaved Caspase-3 (Asp 175) (Cell Signaling
#9661, Danvers, MA, USA), Bcl-2 (D17C4) (Cell Signaling #3498, Danvers, MA, USA), β-Actin (13E5) (Cell Signaling #4970,
Danvers, MA, USA). Secondary antibodies included ones conjugated to Alexa Flour 488 (Abcam, Cambridge, UK), Alexa Flour 647
(Abcam, Cambridge, UK) Alexa Flour 594 (Abcam, Cambridge, UK) for horseradish peroxidase (HRP)-conjugated anti-rabbit antibodies
(Sigma-Aldrich, St. Louis, MO, USA) for 37 chemilluminescence immunoblotting. Antibodies for flow cytometry included APC/fire 750
anti-mouse CD8a (BioLegend #100766), BUV395 Rat Anti-mouse Cd45 (BD Horizon #564279), Purified Rat Anti-Mouse CD16/CD32 Mouse Fc
Block (BD Pharmingen #553142), PerCP-Cy Rat Anti-mouse CD4 (BD Pharmingen #550394), and Anti-Mouse FITC TCR beta (Invitrogen
#1939141)
All antibodies were used according to the manufacture’s recommendation.

Hsu J., Krishnan A., Lee S.A., Dodd-o J.M., Kim B.S., Illei P., Yarnoff K., Hamad A.A., Rabb H, & Bush E.L. (2019). CD3+CD4−CD8− Double-Negative αβ T cells Attenuate Lung Ischemia-Reperfusion Injury. The Journal of thoracic and cardiovascular surgery, 161(1), e81-e90.

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