Anti bax antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-Bax antibody is a laboratory reagent used to detect the Bax protein. Bax is a pro-apoptotic member of the Bcl-2 protein family that plays a role in the regulation of programmed cell death. The Anti-Bax antibody can be used in various techniques, such as Western blotting, immunohistochemistry, and immunofluorescence, to analyze the expression and localization of the Bax protein in biological samples.

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Anti-Bax (288 citations) Bax antibody (18 citations) Mouse anti-Bax antibody (6 citations) Rabbit polyclonal anti-Bax antibody (4 citations) Rabbit anti-Bax antibody (3 citations) Anti-Bax antibody (sc-20067, (2 citations) Anti-BAX monoclonal antibody (2 citations) Goat anti-rat Bax polyclonal antibody (2 citations)

29 protocols using anti bax antibody

Protein Expression Analysis Protocol

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Anti-CHOP, anti-caspase-12, anti-caspase-9, anti-caspase-3, anti-caspase-3 cleaved, anti-Bip, anti-p-eIF2α, anti-p-JNK, anti-p-ERK, anti-p-p38, anti-IRE-1α, anti-Cyt C, and anti-COX IV antibodies were purchased from Cell Signaling Technology; anti-α-tubulin antibodies were acquired from Biyuntian (Nanjing, China); anti-Ag85 and anti-GroEL1 were obtained from Abcam (Cambridge, UK); and anti-BAX antibodies were from Santa Cruz Biotechnology. Goat anti-mouse-IgG-HRP (Proteintech, Wuhan, China) and goat anti-rabbit-IgG-HRP (Proteintech) were used as secondary antibodies. Western blots were detected using the Amersham^TM ECL^TM Prime western blotting detection reagent (GE Healthcare, Buckinghamshire, UK) and the ChemiScope 3400 mini imaging system (Clinx Science Instruments, Shanghai, China). The results shown are representative of three independent experiments.

Guo Q., Bi J., Wang H, & Zhang X. (2021). Mycobacterium tuberculosis ESX-1-secreted substrate protein EspC promotes mycobacterial survival through endoplasmic reticulum stress-mediated apoptosis. Emerging Microbes & Infections, 10(1), 19-36.

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Western Blot Analysis of Cell Signaling

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The cell lysates (20 µg) underwent electrophoresis and were transferred to the nitrocellulose membrane. The latter were blocked in 5% of non-fat milk in PBS 0.1% Tween-20, then incubated in the presence of mouse monoclonal anti-β actin antibody (antibody dilution 1:5000) (A5316 Sigma, St. Loius, MO, USA), rabbit polyclonal anti-Nitric Oxide Synthase-2 (NOS-2) antibodies (antibody dilution 1:200) (sc-651 purchased by Santa Cruz biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-Matrix Metalloproteinases-9 (MMP-9), and anti-Bax antibodies (antibodies dilution 1:200) (sc-5302 and sc-7480, respectively, both purchased by Santa Cruz biotechnology, Santa Cruz, CA, USA), goat polyclonal anti-caspase-3 antibody (antibody dilution 1:200) (sc-1225 purchased by Santa Cruz biotechnology, Santa Cruz, CA, USA). Samples were then probed with specific enzyme conjugated IgG horseradish peroxidase. Immunoreactive bands were revealed by ECL system (Amersham Int., Buckunghamshire, UK) and underwent densitometric analysis. Values obtained from densitometry, expressed as Integrated Optical Intensity (IOI), were evaluated with a CHEMIDOC XRS system through the QuantiOne 1-D analysis software (BIORAD, Richmond, CA, USA). Data were normalized with densitometric values derived from β-actin loading control.

Marconi G.D., Carradori S., Ricci A., Guglielmi P., Cataldi A, & Zara S. (2019). Kinesin Eg5 Targeting Inhibitors as a New Strategy for Gastric Adenocarcinoma Treatment. Molecules, 24(21), 3948.

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Hippocampal Bcl-2 and Bax Expression

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Western blotting for Bcl-2 and Bax was performed according to the previously described method (Kim et al., 2010 (link)). The hippocampal tissues were dissected and collected, and then were immediately frozen at −70°C. The right hemisphere were homogenized on ice, and lysed in a lysis buffer containing 50-mM N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (pH, 7.5), 150-mM NaCl, 10% glycerol, 1% Triton X-100, 1-mM phenylmethylsulfonyl fluoride, 1-mM ethylene glycol tetraacetic acid, 1.5-mM MgCl₂·6H₂O, 1-mM sodium orthovanadate, and 100-mM sodium fluoride. Protein content was measured using a Bio-Rad colorimetric protein assay kit (Hercules, CA, USA). Protein samples (30 μg) were separated on sodium dodecyl sulfate-polyacrylamide gel and transferred onto a nitrocellulose membrane. The membranes were incubated with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 and then incubated overnight at 4°C with the following primary antibodies: mouse anti-β-actin, anti-Bcl-2, and anti-Bax antibodies (1:1,000; Santa Cruz Biotechnology). Subsequently, membranes were incubated for 1 hr with attempt secondary antibodies (1:2,000; Vector Laboratories), and band detection was performed using the enhanced chemiluminescence detection kit (Santa Cruz Biotechnology).

Roh J.H., Ko I.G., Kim S.E., Lee J.M., Ji E.S., Kim J.H., Chang H.K., Lee S.K, & Kim K.H. (2016). Treadmill exercise ameliorates intracerebral hemorrhage-induced depression in rats. Journal of Exercise Rehabilitation, 12(4), 299-307.

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Chlorpyrifos-Induced Neurodegeneration: Mechanisms and Interventions

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Chlorpyrifos (CPF, C₉H₁₁CI₃NO₃PS) was received from Chem Service, Inc., West Chester, PA (cat # S-11459A1). DMSO, glutathione S-transferase and, BHT (3,5-di-tert-butyl-4-hydroxytoluene) were purchased from Sigma (St. Louis, MO, USA). z-VAD-FMK, Z-DEVD-FMK, and Ac-LEHD-AMC were obtained from Cayman chemicals (Ann Arbor, Michigan). Chloromethyl-2’,7’-dichlorofluorescein diacetate (CM-H2DCFDA), JC-1 dye, RPMI 1640, minimal essential medium (MEM), fetal bovine serum, L-glutamine, penicillin and streptomycin were purchased from Invitrogen (Gaithersburg, MD). Antibodies against p-Y701 STAT1, p-S727 STAT1, STAT1, PARP, caspase-3, LC3B, p62 and beclinl were from Cell Signaling Technology (Danvers, MA); anti-NOX-1 and anti-alpha-synuclein (phospho S129) antibodies were purchased from Abeam (Cambridge, MA); and anti-PKCδ, anti-tubulin, anti-Bcl-2, and anti-Bax antibodies were purchased from Santa Cruz Biotech (Dallas, TX). Rat/Mouse Cytochrome c Quantikine ELISA kit was purchased for R&D Systems (Minneapolis, MN). Halt protease and phosphatase inhibitor cocktail (100 x) and SYTOX green dye were purchased from Thermo Fisher Scientific (Waltham, MA). The Cell Death Detection ELSA^plus kit was purchased from Roche Molecular Biochemicals (Indianapolis, IN). N-acetyl cysteine (NAC) and TH antibody were purchased from Calbiochem/EMD Biosciences (Gibbstown, NJ).

Singh N., Lawana V., Luo J., Phong P., Abdalla A., Palanisamy B., Rokad D., Sarkar S., Jin H., Anantharam V., Kanthasamy A.G, & Kanthasamy A. (2018). Organophosphate Pesticide Chlorpyrifos Impairs STAT1 Signaling to Induce Dopaminergic Neurotoxicity: Implications for mitochondria mediated oxidative stress signaling events. Neurobiology of disease, 117, 82-113.

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Evaluating Apoptosis Regulators After Art Treatment

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To determine the expression of Bcl-2 and Bax proteins after art treatment, the harvested cells were fixed overnight in 70% ice-cold ethanol. After washing with ice-cold PBS, the cells were incubated with anti-Bcl-2 (1:100; cat. no. sc-7382; Santa Cruz Biotechnology, Inc.) and anti-Bax antibodies (1:100; cat. no. sc-20067; Santa Cruz Biotechnology, inc.) for 30 min in the dark at room temperature. Subsequently, the cells were incubated with igG-FiTc antibody (1:100; cat. no. 115-095-003; Jackson immunoresearch laboratories, inc.) for 30 min in the dark at room temperature. The stained cells were analyzed using the FC500 flow cytometer, with the mean fluorescence intensity representing the expression of Bcl-2 and Bax proteins.
Statistical analysis. Statistical analysis was performed using the SPSS v21 software (iBM corp.). Values are expressed as the mean ± standard deviation. Multiple groups were compared using one-way analysis of variance followed by Tukey's test. P<0.05 was considered to indicate a statistically significant difference.

Zhao Y., Liu J, & Liu L. (2020). Artesunate inhibits lung cancer cells via regulation of mitochondrial membrane potential and induction of apoptosis. Molecular medicine reports, 22(4).

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DMEM-FBS-Based Cell Culture Protocol

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Dulbecco’s Modified Eagle’s Medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco Lab. (Grand Island, NY, USA). The penicillin-streptomycin solution (100 ×) and RIPA buffer were from Solarbio (Beijing, China). The ECL kit was from Pierce Chemical Co (Rockford, IL, USA). Lipofectamine 2000, TRIzol reagent, Maxima SYBR Green/ROX qPCR Master Mix, and BCA Protein Assay kit were from Thermo Fisher Scientific (Waltham, MA, USA). The M-MLV Reverse Transcriptase was from Promega (Fitchburg, WI, USA). The anti-TRIM55 antibody was from Sigma (Shanghai, China). The anti-DUSP1, cleaved caspase-3, cleaved PARP, and anti-ubiquitin antibodies were from Abcam (Cambridge, MA, USA). The anti-Bcl-2 and anti-Bax antibodies were from Santa Cruz Biotechnology (Dallas, TX, USA). The anti-p-JNK1/2, anti-JNK1/2, and anti-GAPDH antibodies were from Cell Signaling Technology (Danvers, MA, USA). The annexin V-FITC Apoptosis Detection Kit and Horseradish peroxidase-conjugated (HRP)-labeled Goat Anti-Mouse, Donkey Anti-Goat, and Goat Anti-Rabbit IgG secondary antibodies were purchased from Beyotime Biotechnology (Shanghai, China),

Tan J., Shen J., Zhu H., Gong Y., Zhu H., Li J., Lin S., Wu G, & Sun T. (2020). miR-378a-3p inhibits ischemia/reperfusion-induced apoptosis in H9C2 cardiomyocytes by targeting TRIM55 via the DUSP1-JNK1/2 signaling pathway. Aging (Albany NY), 12(10), 8939-8952.

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Neuroprotective Effects of Sodium Tanshinone IIA Sulfonate

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Sodium Tanshinone IIA sulfonate (99.5%) was obtained from Loki's Pharmaceuticals (Beijing, China). Scopolamine hydrobromide injection (Guangzhou Baiyun Mountain Mingxing Pharmaceutical Co., Ltd., Guangzhou, China) was purchased from Guangzhou Pharmaceuticals Corporation (Guangzhou, China). Kits used for detection of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), choline acetyltransferase (ChAT), and acetylcholinesterase (AChE) were purchased from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Primary antibodies (Bcl-2, Caspase-3) were purchased from Cell Signaling Technology, Inc. Anti-Bax antibody was purchased from Santa Cruz Biotechnology, Inc. Anti-β-actin was purchased from Sigma-Aldrich. Secondary antibodies (horseradish peroxidase conjugated anti-rabbit IgG and anti-mouse IgG) were purchased from Cell Signaling Technology, Inc. All other reagents were of the highest grade available commercially.

Xu Q.Q., Xu Y.J., Yang C., Tang Y., Li L., Cai H.B., Hou B.N., Chen H.F., Wang Q., Shi X.G, & Zhang S.J. (2016). Sodium Tanshinone IIA Sulfonate Attenuates Scopolamine-Induced Cognitive Dysfunctions via Improving Cholinergic System. BioMed Research International, 2016, 9852536.

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Western Blot Analysis of Hippocampal Proteins

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Western blot was done, as mentioned in the below (Ji et al., 2020 (link)). The hippocampal tissues were homogenized using lysis buffer consisting of 1 mM EGTA, 1 mM PMSF, 1 mM Na₂VO₄, 1.5 mM MgCl₂·6H₂O, 50 mM Tris-HCl (pH, 8.0), 100 mM NaF, 150 mM NaCl, 1% Triton X-100, 10% glycerol, and then centrifuged at 50,000 rpm for 1 hr. Anti-β-actin antibody (1:2,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-BDNF (1:1,000), anti-TrkB (1:1,000), anti-Bax antibody (1:1,000; Santa Cruz Biotechnology), and anti-Bcl-2 antibody (1:1,000; Santa Cruz Biotechnology) were used as the primary antibodies. Horseradish peroxidase-conjugated anti-mouse antibody for β-actin, Bax, and Bcl-2 (1:3,000; Amersham Pharmacia Biothech GmbH, Freiburg, Germany), and horseradish peroxidase-conjugated anti-rabbit antibody for BDNF and TrkB (1:5,000; Vector Laboratories), were used as the secondary antibodies. In addition to the membrane transfer performed at 4°C, all other steps were performed at room temperature. Band detection was done by enhanced chemiluminescence detection kit (Santa Cruz Biotechnology).

Kim T.W., Ko Y.J., Youn K.H., Hwang B.G., Bang H.S, & Lee S.J. (2021). Treadmill exercise improves spatial learning ability by increasing cell proliferation in offspring born to maternal rats receiving stress during pregnancy. Journal of Exercise Rehabilitation, 17(2), 88-95.

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Neuroprotective Effects of Phytochemical Compounds

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Ginsenoside Rb1, Ginsenoside Rg1, Osthole, Imperatorin, Paeoniflorin, Paeonolum, Oleanic acid and 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside were purchased from the National Institutes for Food and Drug Control (Beijing, China). Acetonitrile [high performance liquid chromatography (HPLC) grade] was bought from Honeywell International Inc. (Burdick & Jackson, Muskegon, MI, USA). SCOP hydrobromide injection (Guangzhou Baiyun mountain Mingxing Pharmaceutical Co., Ltd., Guangzhou, China) was purchased from Guangzhou Pharmaceuticals Corporation (Guangzhou, China). Acricept (Henan Joyline & Joysun Pharmaceutical Stock Co., Ltd., Zhengzhou, China) was dissolved in 0.9% physiological saline. Kits used for determination of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) were purchased from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Primary antibodies (Bcl-2, caspase-3 and β-actin) were obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA). Anti-Bax antibody was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Secondary antibodies (horseradish peroxidase-conjugated anti-rabbit IgG and anti-mouse IgG) were purchased from Cell Signaling Technology, Inc. Other reagents were of AR grade.

HOU X.Q., WU D.W., ZHANG C.X., YAN R., YANG C., RONG C.P., ZHANG L., CHANG X., SU R.Y., ZHANG S.J., HE W.Q., QU Z., LI S., SU Z.R., CHEN Y.B., WANG Q, & FANG S.H. (2014). Bushen-Yizhi formula ameliorates cognition deficits and attenuates oxidative stress-related neuronal apoptosis in scopolamine-induced senescence in mice. International Journal of Molecular Medicine, 34(2), 429-439.

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Bax-mediated Liposome Permeabilization

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Liposomes were generated by the extrusion method as described21 (link) and incubated with Bax (1.8 μM) and cBid (1.26 μM) for 2 h at 37 °C to be permeabilized. After the incubation, the liposomes were floated-up by high-speed centrifugation and collected with microfilters with 0.1 μm pore size. The retained membrane fraction was dissolved in 20 mM HEPES, pH 7.4, 150 mM NaCl containing 1.2% CHAPS and fractionated in Superdex 200 (GE) in the same buffer. Bax alone in buffer was also fractionated. Bax was detected by immunoblotting with anti-Bax antibody (Santa Cruz; N20).

Kuwana T., Olson N.H., Kiosses W.B., Peters B, & Newmeyer D.D. (2016). Pro-apoptotic Bax molecules densely populate the edges of membrane pores. Scientific Reports, 6, 27299.

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