BBR and dimethyl sulfoxide (DMSO) were obtained from Sigma Chemicals. Anti-β-actin, anti-Bcl-2, anti-Bax, anti-TF, anti-Ki67, anti -p38, anti-p-p38, anti -JNK, anti-p-JNK and Anti-β-actin were purchased from Abcam. Other chemicals and experimental materials were provided by BioRad.
Anti β actin
Manufactured by Abcam
Sourced in United States, United Kingdom, China, Hong Kong, Germany, Canada, France
Anti-β-actin is a primary antibody used to detect and quantify the β-actin protein in various cell and tissue samples. β-actin is a widely expressed housekeeping gene and its protein product is commonly used as a loading control in Western blot analysis.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (117 mentions)
Bca protein assay kit, by Beyotime (56 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (53 mentions)
Anti bax, by Abcam (51 mentions)
Anti bcl 2, by Abcam (46 mentions)
Anti gapdh, by Abcam (44 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (44 mentions)
Ripa lysis buffer, by Beyotime (39 mentions)
Anti e cadherin, by Abcam (39 mentions)
Anti vimentin, by Abcam (31 mentions)
Ripa buffer, by Beyotime (30 mentions)
Anti cleaved caspase 3, by Abcam (30 mentions)
Ripa buffer, by Thermo Fisher Scientific (29 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (28 mentions)
Anti n cadherin, by Abcam (25 mentions)
Anti flag, by Merck Group (23 mentions)
Anti akt, by Abcam (23 mentions)
Polyvinylidene difluoride membrane, by Merck Group (23 mentions)
Anti β catenin, by Abcam (22 mentions)
Anti caspase 3, by Abcam (20 mentions)
1 185 protocols using anti β actin
1
Investigating Cellular Signaling Pathways
2
Protein Expression Analysis in fMSCs, hGCs, and Ovarian Tissue
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fMSCs, hGCs, and ovarian tissues were harvested for protein extraction. Western blotting was performed as previously described [12 (link)]. The primary antibodies used for fMSCs were anti-Oct4, anti-Nanog, anti-Rex1, and anti-β-Actin, all purchased from Abcam (USA). The primary antibodies used for hGCs and the ovarian tissues were anti-SURVIVIN, anti-BCL2, anti-CASPASE-3, anti-CASPASE-9, anti-MT1, anti-JNK1, anti-PCNA, anti-AMPK, anti-β-Actin, and anti-GAPDH, all purchased from Abcam (USA).
3
Notch Signaling Pathway Analysis
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RPMI-1640, fetal bovine serum (FBS) and penicillin/streptomycin were obtained from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The lentiviral transcriptional reporter vector, pGreenFire1-Notch-EF1-Puro, was purchased from System Biosciences (#121010-003; Palo Alto, CA, USA). The primary antibodies and mouse monoclonal anti-β-actin were purchased from Abcam (Cambridge, MA USA), which included anti-Notch1 (#ab20687; dilution, 1:1,000), anti-hairy and enhancer of split-1 (Hes-1) (#ab194111; dilution, 1:1,000), anti-vimentin (#ab5741; dilution, 1:1,000) anti-E-cadherin (#ab17021; dilution, 1:1,000), anti-N-cadherin (#ab20191; dilution, 1:1,000) and anti-β-actin (#ab8226; dilution, 1:2,000) antibodies.
4
Comprehensive Antibody Panel for Western Blot and IHC
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For western blotting and immunohistochemistry analyses, the following antibodies were purchased from Abcam (Cambridge, UK): anti-H2A.Z (cat no. ab150402; dilution 1:1,000 for western blot analysis and dilution 1:300 for immunohistochemistry), anti-p21/WAF1/Cip1 (cat. no. ab109520), anti-p27/Kip1 (cat. no. ab32034), anti-Skp2 (cat. no. ab124799), anti-cyclinA (cat. no. ab181591) and matrix metalloproteinase (MMP)2 (cat. no. ab37150). The following antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA): anti-cyclin-dependent kinase (CDK) 4 (cat. no. 12790), anti-CDK6 (cat. no. 3136) and anti-CDK2 (cat. no. 2546). Ki67 (cat. no. ab15580; Abcam) was used for the immunohistochemical analysis of mouse tumor tissue. The following antibodies were purchased from Epitomics (Burlingame, CA, USA): anti-β-actin (cat. no. 1854-1), and anti-GAPDH (cat. no. 5632-1). A kit with antibodies targeting EMT-associated proteins (E-cadherin, N-cadherin, Slug, Snail, and Vimentin) was obtained from Cell Signaling Technology, Inc. (cat, no. 9782S). Antibodies, recognizing total (#9665) and cleaved caspase-3 (#9664), caspase-9 (#9508), Bcl-2 homologous antagonist/killer (Bak) (#12105) and B-cell lymphoma 2 (Bcl-2) (#15071) were obtained from Cell Signaling Technology, Inc.
5
Western Blot Protein Analysis
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Cells (2 × 106) were lysed in RIPA lysis buffer (Beyotime Biotechnology) and quantitated using a BCA Protein Assay Kit (Beyotime Biotechnology). The protein liquid was mixed with 5 × loading buffer in a volume ratio of 4 to 1, and placed in a boiling water bath for 10 min to denature. For Western blot analysis, equivalent amounts of protein per sample were electrophoretically resolved on 10% polyacrylamide gels and then transferred onto PVDF membranes (Millipore). Following this, the PVDF membranes with the protein were blocked with blocking buffer (Beyotime Biotechnology), then incubated with the corresponding primary antibodies. After repeated washes, the membranes were incubated with horseradish‐peroxidase‐conjugated anti‐mouse or anti‐rabbit secondary antibody (Cell Signaling, 1:1000 diluted) at room temperature for 1 h. An electro‐chemiluminescence (ECL) system (Thermo Fisher Scientific) was used for the detection. Anti‐β‐actin (Epitomics) was used to check for equal loading of protein between wells. The Primary antibodies used for the Western blot are shown in Table 2 .
6
Apoptosis Assay Protocol for Cell Lines
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Calcium ionophore (A23187), benzamidine hydrochloride, N-acetyl-Leu-Glu-His-Asp trifluoro methylcoumarin (AC-LEHD-FMC), acetyl-Asp-Glu-Val-Asp-7-amido-4-methylcoumarin (AC-DEVD-AMC), sodium orthovanadate, dimethyl sulfoxide (DMSO), fluorescein isothiocyanate (FITC)-labeled annexin V, 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetylester (CM-H2DCFDA), CHAPS, rhodamine 123, leupeptin hydrochoride, N-(2-Hydroxyethyl) piperazine-N′-ethanesulfonic acid (HEPES), fura-2/AM, monoclonal anti-phosphotyrosine antibody, acridine orange 10-nonyl bromide (NAO) and dithiothreitol (DTT) were from Sigma Chemicals, St. Louis (USA). Monoclonal anti-cytochrome c antibody and anti-β-actin were from Epitomics Burlingame, CA (USA). Anti-Caspase-3 antibody was from Santa Cruz Biotechnology, Inc. Texas (USA). Collagen type-I was from Chrono-log Corporation, Pennsylvania (USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 1, 1-diphenyl-2-picrylhdrazyl (DPPH) were from HiMedia Laboratories, Mumbai (India). Lactate dehydrogenase (LDH) kit was from AGAPPE diagnostics Ltd., Kerala (India). γ-glutamyl p-nitroanilide and glycylglycine were from Sisco Research laboratories Pvt Ltd., Mumbai (India). All other reagents were of analytical grade.
7
Paclitaxel and Cisplatin Cytotoxicity Assay
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Paclitaxel was purchased from Bristol-Myers Squibb Pharmaceutical Ltd. Cisplatin (#15663-27-1) was purchased from MedChem Express. Antibodies used for Western blotting included anti-SOX2 (#3579, CST), anti-OCT4 (#19857, Abcam), anti-β-actin (#1879-1, Epitomics), anti-mouse HRP (#7076, Cell Signaling Technology), and anti-rabbit HRP (#7074, Cell Signaling Technology).
8
Protein Interaction Assay with Compounds
9
Western Blot Analysis of FLI-1 Protein
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Cells were lysed in RIPA buffer. Cellular proteins were collected and subjected to 10% SDS-PAGE, and transferred onto PVDF membranes (Amersham Pharmacia Biotech, Piscataway, NJ). The membranes were then incubated with specific primary antibodies. Anti-FLI-1 was purchased from Santa Cruz Biotechnology, Inc. Anti-β-ACTIN was purchased from Epitomics, Inc. This was followed by incubation with horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG antibodies from Santa Cruz Biotechnology, and antigen-antibody complexes were visualized using the Western Bright ECL detection system (Advansta, CA).
10
Protein Expression Analysis by Western Blotting
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The proteins were then extracted based on the manufacturer's instructions. Equivalent amounts of protein per sample were electrophoretically resolved on 10% polyacrylamide gels and transferred onto 0.2 mm nitrocellulose membranes. Proteins were probed overnight with primary antibodies against AQP3 (1:1000; ab125219, Abcam), beclin-1 (1:1000; GTX 31,722, GeneTex), LC3B (1:1000; CST 3868, Cell Signaling Technology, Danvers, MA, USA), actin (1:5000, MAB1501, Millipore, Burlington, MA, USA). The nitrocellulose membranes were then incubated with an appropriate horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature, and the immunoreactivity was observed by enhanced chemiluminescence detection, a semiquantitative assay through blot dosimetry. Anti-β-actin (Epitomics, Cambridge, MA, USA) was used to check for equal loading of protein between wells25 (link).
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