Bcl 2

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Sourced in United States, China

Bcl-2 is a protein that plays a key role in the regulation of apoptosis, or programmed cell death. It functions as an anti-apoptotic protein, helping to prevent cell death and promote cell survival. Bcl-2 is a commonly used marker in various cell biology and molecular biology applications, including immunohistochemistry, flow cytometry, and Western blotting.

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Lab products found in correlation

Bcl2 associated x (bax), by GeneTex (11 mentions) Gapdh, by GeneTex (7 mentions) Caspase 3, by GeneTex (5 mentions) Pvdf membrane, by Merck Group (4 mentions) Cleaved caspase 3, by Cell Signaling Technology (3 mentions) Caspase 3, by Cell Signaling Technology (3 mentions) Caspase 9, by Cell Signaling Technology (3 mentions) Cytochrome c, by GeneTex (3 mentions) Cleaved caspase 3, by GeneTex (2 mentions) Gapdh, by Abcam (2 mentions) Bcl2 associated x (bax), by Abcam (2 mentions) Cleaved caspase 9, by Cell Signaling Technology (2 mentions) Bcl xl, by GeneTex (2 mentions) Caspase 8, by GeneTex (2 mentions) Beclin 1, by Cell Signaling Technology (2 mentions) N hydroxysuccinimide nhs, by Merck Group (2 mentions) β actin, by Merck Group (2 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Quantity one analysis software, by Bio-Rad (1 mentions) Nuclear separation kit, by NJJCBIO (1 mentions)

Close spelling variants of this product

Anti-Bcl-2 (10 citations) Antibody for Bcl-2 (GTX100064) (2 citations) Mouse monoclonal anti-human BCL-2 (124; (2 citations)

22 protocols using bcl 2

Cytoplasmic and Nuclear Protein Isolation

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The cytoplasm and nuclei in LV tissue and RAW264.7 macrophages were isolated using a nuclear separation kit (Njjcbio) according to a previously described protocol [19 (link)]. Briefly, homogenate was collected from LV tissue and cells that were lysed in lysis buffer and further centrifuged at 1000 × g for 10 minutes. The pellets in the bottoms of the tubes contained the nuclei, and the supernatant contained the cytoplasm.
After the cytoplasm, nuclei, LV tissue, and HL-1 cardiomyocytes were, respectively, lysed in RIPA lysis buffer containing 10% protease inhibitors and 10% phosphatase inhibitors, the total protein was obtained and quantitated using a BCA Protein Assay Kit. After separation by electrophoresis on 10% SDS polyacrylamide gels, the proteins were transferred to PVDF membranes. Then, the protein expression of IL-16 (Abcam), Bax, Bcl-2, cleaved caspase-3, caspase-3, and GAPDH (all three from GeneTex) in total LV tissue, the protein expression of p-p65 and GAPDH (both from Abcam) in the cytoplasm, the protein expression of p-p65 and PCNA (GeneTex) in the nuclei, and the protein expression of Bax, Bcl-2, cleaved caspase-3, caspase-3, and GAPDH in HL-1 cardiomyocytes were measured using primary antibodies as indicated in parentheses. After further incubation with the secondary antibodies, the target proteins were detected and analyzed.

Zhang J., Yang Z., Liang Z., Wang M., Hu C., Chang C., Shi L., Ji Q, & Liu L. (2021). Anti-Interleukin-16-Neutralizing Antibody Attenuates Cardiac Inflammation and Protects against Cardiac Injury in Doxorubicin-Treated Mice. Mediators of Inflammation, 2021, 6611085.

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Aortic and Macrophage Gene Expression Analysis

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Western blot and quantitative RT–qPCR analyses were performed according to our previous studies [12 (link)–14 (link)]. In this study, the aortic expression levels of IL-22, Bax, Bcl2, and GAPDH (all from GeneTex) were determined by western blot analysis, and the mRNA levels of IL-22, iNOS, CD38, CD80, and CD86 in macrophages and of Bax and Bcl2 were measured by RT–qPCR. The primers are provided in Supplementary Material Table S1.

Wang Y., Li J., Xu Y., Liao S., Song J., Xu Z., Wei W, & Zhu S. (2022). Interleukin-22 Deficiency Reduces Angiotensin II-Induced Aortic Dissection and Abdominal Aortic Aneurysm in ApoE-/- Mice. Oxidative Medicine and Cellular Longevity, 2022, 7555492.

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Western Blot Analysis of Neural Markers

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The cells were washed twice with ice-cold phosphate-buffered saline (PBS) and directly lysed in Laemmli buffer. The lysate was sonicated, boiled for 5 min and centrifuged at 16,000×g for 10 min at 4°C. The supernatant was recovered as total cell lysate, aliquoted and stored at −80°C. Equal amounts of protein (10 µg) were separated through 8% SDS-PAGE and electro-transferred onto 0.45 µm polyvinylidene difluoride membranes (Millipore, Bedford, USA). Following transfer, the membranes were blocked with a solution of 0.1% Tween 20/TBS (TBS/T) containing 5% non-fat milk for 1 h at room temperature and subsequently incubated overnight at 4°C with monoclonal mouse anti-human CGRP (Santa Cruz Biotechnology, Santa Cruz, CA, USA, final dilution 1∶400), Bcl-2 (GeneTex, USA, final dilution 1∶300), Wnt-1, Wnt-3a, Wnt-5a, Wnt-7 and β-catenin (Sigma, St. Louis, MO, USA, final dilution 1∶400) antibodies or rabbit polyclonal anti-human nestin, MAP2, RIP, and GFAP antibodies (Sigma, St. Louis, MO, USA, final dilution 1∶500). The bands were visualized using nitroblue tetrazolium/5-bromo-4-chloro-3-indolyl-phosphate. GAPDH served as an endogenous control. For densitometric analyses, the blots were scanned and quantified using Quantity One analysis software (Bio- Rad, Hercules, CA, USA). The results were expressed as a percentage of GAPDH immunoreactivity.

Yang Q., Du X., Fang Z., Xiong W., Li G., Liao H., Xiao J., Wang G, & Li F. (2014). Effect of Calcitonin Gene-Related Peptide on the Neurogenesis of Rat Adipose-Derived Stem Cells In Vitro. PLoS ONE, 9(1), e86334.

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Regulation of mTOR Signaling Pathway

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Primary antibodies specific to mTOR were purchased from Abcam (Cambridge, MA, USA). Akt, phospho-mTOR (Ser2448), phospho-Akt (Ser473), phospho-AMPKα (Thr172), AMPKα, cleaved caspase 3, and cleaved caspase 9 (Cell Signaling; Danvers, MA, USA); Bcl-XL, Bcl-2, Bax, Bak, poly ADP-ribose polymerase (PARP), p62, microtubule-associated proteins 1A/1B light chain 3B (LC3B), and GAPDH (Genetex; Irvine, CA, USA); and cyclin-dependent kinase 1 (CDK1) and cyclin B1 (Merck Millipore; Burlington, MA, USA). Anti-rabbit polyclonal and anti-mouse monoclonal antibodies were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). ROS scavengers N-acetylcysteine (NAC) and H₂O₂-scavenging enzyme catalase were purchased from Merck Millipore (Burlington, MA, USA). Pan-caspase inhibitor z-VAD-FMK was purchased from R&D Systems (Minneapolis, MN, USA). Naringenin (N5893) and all other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Chang T.M., Chi M.C., Chiang Y.C., Lin C.M., Fang M.L., Lee C.W., Liu J.F, & Kou Y.R. (2024). Promotion of ROS-mediated apoptosis, G2/M arrest, and autophagy by naringenin in non-small cell lung cancer. International Journal of Biological Sciences, 20(3), 1093-1109.

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Western Blot Analysis of Protein Expression

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Protein samples were separated by 8%, 10%, or 12% sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE) and then transferred onto PVDF membranes (EMD Millipore, Burlington, MA, USA). After being blocked with 5% non-fat milk, the membranes were incubated with the following primary antibodies at 4°C overnight: α-tubulin (GeneTex, Irvine, CA, USA), collagen I (GeneTex), α-SMA (Proteintech, Chicago, IL, USA), TGF-β (Cell signaling technology, Danvers, MA, USA), TNF-α (GeneTex), NLRP3 (Proteintech), caspase-1 (Proteintech), IL-18 (Proteintech), IL-1β (Proteintech), caspase-3 (GeneTex), caspase-8 (GeneTex), caspase-9 (GeneTex), and BCL-2 (GeneTex). Membranes were then incubated with HRP-conjugated mouse anti-IgG (EMD Millipore) or HRP-conjugated rabbit anti-IgG (EMD Millipore) secondary antibodies for 1 h. Membranes were developed using ECL detection reagent (EMD Millipore). Relative protein levels were quantified using Image J (Version 1.46, National Institute of Health, Bethesda, MD, USA), and protein densitometry were expressed relative to that of α-tubulin.

Lam H.Y., Liang T.R, & Peng S.Y. (2021). Ameliorative effects of Schisandrin B on Schistosoma mansoni-induced hepatic fibrosis in vivo. PLoS Neglected Tropical Diseases, 15(6), e0009554.

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Western Blot Analysis of HCMEC Proteins

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Total proteins that extracted from HCMECs utilizing RIPA buffer (JRDUN Biotechnology) were ascertained with the application of BCA protein assay kit (CoWin, Beijing, China). Following, PVDF membranes were to shift protein samples that resolved by 10% SDS-PAGE. Non-specific interaction was ensured after the membranes were incubated with 5% non-fat milk. Afterwards, primary antibodies as well as goat anti-rabbit HRP antibody (cat. no. ab205718; 1/2000; Abcam) were respectively probed with the membranes overnight at 4°C and for 1 h. The blots were visualized by the ECL western blotting system (Amersham; Cytiva) and analysed by Image Quant LAS 500 (GE Healthcare) [36] (link). CTRP3 (cat. no. GTX85505; 1:1000; GeneTex), B cell lymphoma-2 (Bcl-2; cat. no. ab32124; 1/1000; Abcam), Bcl-2-associated X (Bax; cat. no. ab32503; 1/1000; Abcam), cleaved caspase 3 (cat. no. ab2302; 1/1000; Abcam), cytochrome c (cyto-C; cat. no. ab133504; 1/5000; Abcam), zonula occludens-1 (ZO-1; cat. no. ab216880; 1/5000; Abcam), Occludin (cat. no. ab216327; 1/1000; Abcam), VE-cadherin (cat. no. ab33168; 1/1000; Abcam), Claudin-5 (cat. no. ab131259; 1/1000; Abcam), FOXO6 (cat. no. 19122-1-AP; 1/500; Proteintech), SIRT1 (cat. no. ab189494; 1/1000; Abcam), Nrf2 (cat. no. ab62352; 1/1000; Abcam), GAPDH (cat. no. ab9485; 1/2500; Abcam) antibodies were utilized here.

Zheng S., Wang Y., Guo W, & Tan H. (2024). FOXO6 transcription inhibition of CTRP3 promotes OGD/R-triggered cardiac microvascular endothelial barrier disruption via SIRT1/Nrf2 signalling. Folia morphologica, 83(1).

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Cytotoxicity Assay of Complex 5 in MDAMB231 Cells

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MDAMB231 cells were seeded into culture flask. After incubation for 24 h, the cells were treated with complex 5 (10 μM–120 μM) for 24 h. Then the cell lysate was obtained and the protein concentration was measured. The same amount of protein (20 μg) was separated from 10% SDS-PAGE gel and transferred to polyvinylidene fluoride (PVDF) membrane (MilliporeCorporation, United States). The imprinting was sealed in a closed buffer at room temperature for 2 h, and then incubated with the diluted solution (1:500–1:1000) of Caspase-3 (abcam, 96625), Bax (genetex, 109683), Bcl-2 (genetex, 100629), HO-1 (abcam, 68477). The antibody (US) blocked the buffer overnight at 4°C. Then the membranes were incubated with the appropriate secondary antibody (1:5000–1:10000 dilution), and GAPDH (immunoway, B1501) was used as a loading control. The protein bands were visualized by X-ray dark box (11–14 of Shantou Yongtai Medical Instrument Co., Ltd.).

Liu H., Chen Y., Cui F., Liao Y, & Wang X. (2022). Synthesis, docking studies, biological activity of carbon monoxide release molecules based on coumarin derivatives. Frontiers in Chemistry, 10, 996079.

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Western Blot Analysis of Signaling Proteins

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Cells were lysed in ice-cold RIPA buffer (50 mM of Tris-HCl pH 7.4, 150 mM of NaCl, 1% Triton X-100, 0.25% deoxycholate, 1.5 mM of MgCl₂, 1 mM of EGTA, 1 mM of phenylmethylsulfonyl fluoride, 10 mM of ZnAF, 10 mM of pervanadate, 10 µg/ml of leupeptin and 10 µg/ml of aprotinin) and incubated for 30 min. The protein concentration was determined using the BCA method. The cell lysate was heated at 100°C for 10 min after being mixed with sample loading buffer. The protein samples were equally loaded on 10% SDS-PAGE, and transferred onto nitrocellulose membranes. After being blocked with 5% non-fat milk in Tris-buffer, the membrane strips were incubated with a primary antibody overnight at 4°C, followed by Alex Fluor 680/790-labeled goat anti-rabbit or goat anti-mouse IgG (Li-COR Biosciences, Lincoln, NE, USA). The strips were visualized using the LI-COR Odyssey Infrared Imaging System (Li-COR Biosciences).
The primary antibodies were as follows: p38, phospho-p38, ERK, phospho-ERK, JNK and phospho-JNK (1:1,000; Cell Signaling Technology, Danvers, MA, USA); MMP2, VEGF, EGFR, cyclin D and E (1:1,000; Abcam, Cambridge, MA, USA); p65, Bcl2, Snail, c-MYC, SSRP1 and GAPDH (1:1,000; GeneTex, Inc.).

Liao J., Tao X., Ding Q., Liu J., Yang X., Yuan F.E., Yang J.A., Liu B., Xiang G.A, & Chen Q. (2017). SSRP1 silencing inhibits the proliferation and malignancy of human glioma cells via the MAPK signaling pathway. Oncology Reports, 38(5), 2667-2676.

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Western Blot Analysis of Apoptosis Markers

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The cell lysates treated with X.E.E. or X.H.E. at the indicated concentrations for various periods of time were prepared for immunoblotting of LC3, p53, Caspase 3, Bcl2, Bcl2-associated X protein (Genetex, USA). Western blot analysis was performed as previously reported [29 (link)]. The original blots can be referred in the additional file 1.

Huang Y.Y., Chen C.H., Hsu C.H., Kuo T.Y., Liu C.C., Liao A.T, & Lin C.S. (2020). Inhibiting autophagy potentiates the antitumor efficacy of Euphorbia royleana for canine mammary gland tumors. BMC Veterinary Research, 16, 193.

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Protein Extraction and Western Blotting

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Total cellular protein was extracted using a radioimmunoprecipitation assay buffer (50 mM Tris-Cl, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, and 1% sodium deoxycholate) and samples were resolved by using SDS-PAGE analysis. A bicinchoninic acid protein quantitative kit (Thermo Fisher Scientific Inc.) was used for protein concentration determination. Each lane was loaded with protein samples (25 µg) and then resolved by 10% SDS-PAGE gel and transferred onto a PVDF membrane (EMD Millipore, Billerica, MA, USA) and blocked with Tris-buffered saline with 0.1% Tween-20 containing 5% non-fat milk for 1 h at room temperature and then blotted overnight at 4°C with primary antibodies against TP73 (1:1,000; cat. no. N2C1; GeneTex, Inc., Irvine, CA, USA), Bcl-2 (1:1,000; cat. no. ab59348) and Bax (1:1,000; cat. no. ab32503) or GAPDH (1:2,000; cat. no. ab8245; all Abcam, Cambridge, UK), and incubated with HRP-conjugated anti-rabbit IgG antibody (1:2,000; cat. no. 7074; Cell Signaling Technology Inc., Danvers, MA, USA) at room temperature for 1 h. Protein bands were observed using enhanced chemiluminescence (SuperSignal West Pico Chemiluminescent substrate; Thermo Fisher Scientific, Inc.) and then analyzed using ImageJ software (version 1.46; National Institutes of Health, Bethesda, MD, USA).

Zhang X., Zhang M., Wang G., Tian Y, & He X. (2018). Tumor promoter role of miR-647 in gastric cancer via repression of TP73. Molecular Medicine Reports, 18(4), 3744-3750.

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