β actin ab179467

Manufactured by Abcam

Sourced in United States, United Kingdom

β-actin (ab179467) is a recombinant antibody that detects the β-actin protein. β-actin is a ubiquitous and highly conserved cytoskeletal protein that is involved in various cellular processes, including cell motility, structure, and integrity. This antibody can be used for detection and quantification of β-actin expression in various applications.

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Lab products found in correlation

Bio spectrum gel imaging system, by Bio-Rad (1 mentions) Tris buffered saline tween tbst, by Boster Bio (1 mentions) Gapdh ab181602, by Abcam (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Proteinase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Novex gel, by Thermo Fisher Scientific (1 mentions) Ecl substrate, by Thermo Fisher Scientific (1 mentions) Nitrocellulose filter membrane, by Merck Group (1 mentions) Protease inhibitor, by Solarbio (1 mentions) Sample buffer, by Solarbio (1 mentions) P erk1 2, by Cell Signaling Technology (1 mentions) P rps6, by Cell Signaling Technology (1 mentions) Ecl substrate, by Merck Group (1 mentions) Bs 7175r, by Bioss Antibodies (1 mentions) Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions) Pakt s473, by Cell Signaling Technology (1 mentions) P mtor, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Erk1 2, by Cell Signaling Technology (1 mentions) Ripa buffer, by Solarbio (1 mentions)

6 protocols using β actin ab179467

Western Blot Analysis of Exosomal Markers

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The extraction of the total protein was performed via radio-immunoprecipitation assay lysis buffer embodying phenylmethylsulfonyl fluoride (Beyotime Institute of Biotechnology, Shanghai, China). The protein level in the supernatant was detected via the BCA method. Equal volumes of protein (50 mg) were separated via SDS-PAGE (10% gel) and then transferred onto the polyvinylidene fluoride (PVDF) membrane (EMD Millipore). The PVDF membranes were incubated with tris-buffered saline tween (TBST; Boster Biological Technology Co., Ltd.) supplemented with 5% skimmed milk to block the non-specific binding. Next, the membranes were cultured with the primary antibodies (Table 1) at 4°C overnight, together with rabbit anti-rat secondary antibody for 1 h at room temperature. The proteins were developed in enhanced chemiluminescence reagent, and analyzed by BioSpectrum gel imaging system (Bio-Rad Laboratories, Inc.).Table 1

Antibodies for Western Blot Analysis

Antibody	No., Company	Dilution Ratio
Alix	ab117600, Abcam	1: 100
CD63	ab217345, Abcam	1: 100
CD9	ab92726, Abcam	1: 100
CD81	ab79559, Abcam	1: 5000
GAPDH	ab181602, Abcam	1: 50
β-actin	ab179467, Abcam	1: 5000
Cleaved caspase-3	ab2302, Abcam	1:50
Cleaved PARP	ab32064, Abcam	1: 5000
Secondary antibody	ab150117, Abcam	1: 5000

Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PARP, poly (ADP-ribose) polymerase; Abcam, Abcam Inc., Cambridge, MA, USA.

Bu X., Li D., Wang F., Sun Q, & Zhang Z. (2020). Protective Role of Astrocyte-Derived Exosomal microRNA-361 in Cerebral Ischemic-Reperfusion Injury by Regulating the AMPK/mTOR Signaling Pathway and Targeting CTSB. Neuropsychiatric Disease and Treatment, 16, 1863-1877.

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Immunoblotting Analysis of STING and IRF-3 Signaling

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The tumor tissues were homogenized by bead beating in RIPA buffer containing with 1% proteinase inhibitor cocktail (200 μL/20 mg tissue, ThermoFisher, #78429) within 10 min at 4 °C. The lysates were then centrifuged at 15,000 r.p.m. for 20 min at 4 °C, the supernatants were collected and quantified by BCA assay. Samples were dissolved in SDS-PAGE protein loading buffer (5X, Boster, AR1112) and boiled for 10 min, and equal amounts of each sample were applied to 12% Tris-glycine gel (Novex gel, Invitrogen) and subjected to electrophoresis, which was subsequently transferred to a PDVF (polyvinylidene difluoride) membrane and blocked in 5% non-fat milk blocking buffer for 1 h at room temperature. The membranes were incubation with the primary antibody overnight at 4 °C (STING (D1V5L) mAb, #50494, dilution 1/1000; Phospho-STING (Ser365) (D8F4W) mAb, #72971, dilution 1/1000; IRF-3 (D83B9) mAb, #4302, dilution 1/1000; Phospho-IRF-3 (Ser396) (D6O1M) mAb, #29047, dilution 1/1000, Cell Signaling; β-Actin, ab179467, dilution 1/1000, Abcam), and then horseradish peroxidase (HRP)-conjugated secondary antibodies (anti-rabbit IgG, HRP-linked antibody, #7074, dilution 1/3000; Cell Signaling). Finally, the membranes were soaked in ECL substrate (Thermo Scientific) and imaged under Azure Biosystems software (v. 1.5.0.0518).

Wang Z., Li W., Park J., Gonzalez K.M., Scott A.J, & Lu J. (2022). Camptothesome Elicits Immunogenic Cell Death to Boost Colorectal Cancer Immune Checkpoint Blockade. Journal of controlled release : official journal of the Controlled Release Society, 349, 929-939.

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Western Blot Analysis of Lung Cancer

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Lung cancer cells or tumor tissues were lysed and sonicated in RIPA buffer supplemented with protease inhibitors (Solarbio, Beijing). Protein extracts were boiled in sample buffer (Solarbio), separated by SDS-PAGE and transferred to nitrocellulose filter membranes (Millipore). After blocking in PBS/Tween-20 containing 5% BSA, the membranes were incubated with the primary antibodies. The following antibodies were used: PCNA (#2586), GAPDH (#5174), p-Erk1/2 (#9101), Erk1/2 (#9102), p-AKT (S473, #9271), AKT (#2920), p-RPS6 (#2215), RPS6 (#2217), p-mTOR (#5536) and mTOR (#2983), purchased from Cell Signaling Technologies; MAL2 from Bioss (bs-7175R). β-Actin (ab179467) was purchased from abcam. Signal visualization was detected with ECL substrate (millipore) and a Biotek imaging system. Immunohistochemistry was performed as previously described [35 (link)].

Lian Z., Yan X., Diao Y., Cui D, & Liu H. (2022). T cell differentiation protein 2 facilitates cell proliferation by enhancing mTOR-mediated ribosome biogenesis in non-small cell lung cancer. Discover. Oncology, 13, 26.

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Western Blot Analysis for Protein Markers

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Western blot analysis was conducted as previously described.23 (link) Primary antibodies, including Flotillin-1 (ab41927), Tsg101 (ab125011), CD63 (ab217345), p21 (ab188224), p16^INK4a (ab211542), and β-actin (ab179467) were purchased from Abcam. PNUTS (14171) was purchased from Cell Signaling Technology.

Xia W., Chen H., Chen D., Ye Y., Xie C, & Hou M. (2020). PD-1 inhibitor inducing exosomal miR-34a-5p expression mediates the cross talk between cardiomyocyte and macrophage in immune checkpoint inhibitor–related cardiac dysfunction. Journal for Immunotherapy of Cancer, 8(2), e001293.

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Comprehensive Cell Death Pathway Modulation

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Necrostatin-1 (S8037), ferrostatin-1 (S7243), Z-VAD-FMK (S7023), RSL3 (S8155), erastin (S7242), deferoxamine (S5742), eugenol (S4706), necrostatin-1 (S8037), necrosulfonamide (S8251), and rotenone (S2348) were bought from Selleck Chemicals. Buthionine sulfoximine (BSO, B2515), diethyl butylmalonate (DBM, 112038), NaN3 (S8032), MG-132 (M7449), CHX (C7698), and DMSO (156914) were obtained from Sigma-Aldrich. Antimycin (1397-94-0) was purchased from Santa Cruz Biotechnology. Phosphate-buffered saline (10010023) and Opti MEM medium (51985034) were obtained from Gibco BRL. Primary antibodies against BRD7 (ab56036), phospho-p53 (ab33889), p53 (ab131442), SLC25A28 (ab80467), VDAC (ab14734), Lamin B (ab16048), and β-actin (ab179467) were obtained from Abcam Technology. Anti-mouse IgG (7076) and anti-rabbit IgG (7054) were bought from Cell Signaling Technology.

Zhang Z., Guo M., Shen M., Kong D., Zhang F., Shao J., Tan S., Wang S., Chen A., Cao P, & Zheng S. (2020). The BRD7-P53-SLC25A28 axis regulates ferroptosis in hepatic stellate cells. Redox Biology, 36, 101619.

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Colorectal Cell Culture and Antibody Profiling

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Human colorectal DLD-1 cells and normal colon epithelial NCM460 cells were purchased from the the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China), and cultured in McCoy's 5A and DMEM medium respectively, both supplemented with 10% fetal bovine serum (Gibco, NY, USA), 2 mM glutamine, 100 units/ml streptomycin and penicillin (Invitrogen, Carlsbad, CA). The cells were grown at 37°C in a humidified 5% CO₂ atmosphere. Monoclonal antibodies specific for Ki67 (ab1667), PCNA (ab92552), AhR (83200S), β-catenin (8480S), LEF1 (2230S), TCF4 (2569S), GAPDH (5174S) and β-actin (ab179467) were obtained from Abcam plc. (Cambridge, UK) and Cell Signaling Technology (Beverly, MA, USA).

Zhang L., Ji Q., Chen Q., Wei Z., Liu S., Zhang L., Zhang Y., Li Z., Liu H, & Sui H. (2023). Akkermansia muciniphila inhibits tryptophan metabolism via the AhR/β-catenin signaling pathway to counter the progression of colorectal cancer. International Journal of Biological Sciences, 19(14), 4393-4410.

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