Tom40 rabbit polyclonal antibodies were from Primm Biotech, and were used at a dilution of 1:5000 for detection on Western blots. The antibodies were purified by affinity chromatography and used at a dilution of 1:300 for immunocytochemistry. Tom20 (sc-11415, 1:1000) and Tom22 (J-31, sc-101286, 1:1000) rabbit polyclonal antibodies were from Santa Cruz. Tom20 and Tom22 mouse monoclonal antibodies were from Abcam (ab56783, 1:1000, and ab57523, 1:1000, respectively). GPR75/HSPA9 mouse monoclonal antibodies were from Abcam (ab2799, 1:1000), and VDAC rabbit polyclonal antibodies were from Millipore (AB10527, 1:2000). β-actin mouse monoclonal antibodies were from Cell Signaling (D68A8, 1:5000). Goat polyclonal anti-rabbit IgG antibodies were from Cell Signaling and were used at dilutions of 1:4000 for Tom40 and VDAC and at 1:5000 for Tom20. Horse polyclonal anti-mouse IgG antibodies were from Cell Signaling and were used at a dilution of 1:2000 for Tom22 and HSPA9. Mouse monoclonal anti-HSPA9 antibodies were from abcam, and were used at 1:1000. Complex I (ab109721) and complex IV (ab109909) activity kits were from Abcam.
Mouse monoclonal β actin antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Mouse monoclonal β-actin antibody is a laboratory tool used to detect the presence and quantity of the β-actin protein in biological samples. β-actin is a ubiquitous cytoskeletal protein that is essential for various cellular processes. This antibody can be used in techniques such as Western blotting, immunohistochemistry, and immunofluorescence to identify and quantify β-actin in a wide range of cell and tissue types.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Express one step sybr r green er kit, by Thermo Fisher Scientific (2 mentions)
Polyacrylamide gel, by Lonza (2 mentions)
Ab56783, by Abcam (1 mentions)
Ab2799, by Abcam (1 mentions)
Tom20 sc 11415, by Santa Cruz Biotechnology (1 mentions)
Murine il 4, by Thermo Fisher Scientific (1 mentions)
Cox 2 rabbit polyclonal antibody, by Thermo Fisher Scientific (1 mentions)
Deae dextran hydrochloride, by Merck Group (1 mentions)
Recombinant human il 1β, by R&D Systems (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Wisent (1 mentions)
Phosphate buffered saline (pbs), by Wisent (1 mentions)
Sp600125, by Bio-Techne (1 mentions)
Recombinant human il 13, by Thermo Fisher Scientific (1 mentions)
Sb203580, by Bio-Techne (1 mentions)
Normal mouse igg, by Santa Cruz Biotechnology (1 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions)
15 protocols using mouse monoclonal β actin antibody
1
Mitochondrial Protein Immunodetection
2
Reagents and Antibodies for Cell Culture
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DMEM, RPMI, fetal bovine serum (FBS) and l -glutamine were purchased from Gibco Life Technologies Ltd., Burlington, Ontario, Canada. Penicillin/streptomycin, phosphate buffered saline (PBS) were from Wisent, St. Bruno, Quebec, Canada. Human recombinant IL-1β was product of R&D Systems (Minneapolis, MN, USA), and LPS endotoxin (Escherichia coli, serotype O128:B12) and DEAE-Dextran hydrochloride of Sigma (Oakville, Ontario, Canada), while murine IL-4 was from Peprotech, Quebec, Canada. Antibodies used in this study were: COX-2 rabbit polyclonal antibody from Thermo Fisher Scientific (Burlington, Ontario, Canada), and COX-2 (C-20) goat polyclonal, NFκB p65 (C-20), p50 (H-119) and Lamin A (H-102) rabbit polyclonal antibodies from Santa Cruz (Dallas, TX, USA). ESE-1 rabbit monoclonal antibody was produced in our laboratory in collaboration with Epitomics, Burlingame, CA, USA [22 (link)]. Hsp90 rabbit polyclonal and β-actin mouse monoclonal antibodies were purchased from Cell Signaling Technology (Whitby, Ontario, Canada).
3
Recombinant IL-13 Signaling Pathway Modulation
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Recombinant human IL-13 (Peprotech, Rocky Hill, NJ, USA) was dissolved in distilled water and added to the culture medium at a final concentration of 1, 5, or 10 ng/mL. The antibodies for immunofluorescence and immunohistochemistry staining were used as follows: Anti-IL-13Rα2 mouse monoclonal antibody (Abcam, Cambridge, UK), normal mouse IgG (Santa Cruz Biotechnology, Dallas, TX, USA), and goat anti-mouse IgG conjugated with Alexa Fluor 488 dye (Thermo Fisher Scientific, Waltham, MA, USA). The antibodies for western blotting were used as follows: Anti-ERK1/2, JNK, p38 MAPK, phospho-ERK1/2 (Thr202/Tyr204), phospho-JNK (Thr183/Tyr185), and phospho-p38 MAPK (Thr180/Tyr182) rabbit monoclonal antibodies and β-actin mouse monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA) as primary antibodies, and anti-mouse IgG and anti-rabbit IgG HRP-linked antibody (Cell Signaling Technology) as secondary antibodies. Signal transduction inhibitor U0126 (ERK1/2 inhibitor) was purchased from Cell Signaling Technology. SP600125 (JNK inhibitor) and SB203580 (p38 inhibitor) were obtained from Tocris Bioscience (Bristol, UK).
4
Antibody Use for Cell Signaling
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The BCKDK (E-12) (1:1000, sc-374425) antibodies were purchased from Santa Cruz Technology. Flag tag and talin-1 (#4021) antibodies were purchased from Sigma-Aldrich. All the following antibodies were purchased from Cell Signaling Technology: ERK1/2 (#4695), p-ERK1/2 (Thr202/Tyr204) (#4649), MEK1/2 (8727s), p-MEK1/2 (S221) (2338s), β1-integrin (D6S1W), TRIM21 (#92043), AKT (#4060), p-AKT (S473) (#13038), HA tag antibody (#3724), β-actin mouse monoclonal antibody (#3700), Anti-rabbit IgG (Alexa Fluor#488 Conjugate) (4412), Anti-mouse IgG (Alexa Fluor#594 Conjugate) (8890), secondary antibody against mouse and rabbit. The FAK (A11131), pY397-FAK (AP0302), talin-2 (A19810), and BCAT2 (A7426) antibodies were purchased from ABclonal. The vinculin (ab129002), BCKDHA (ab305168), p-BCKDHA (S293) (ab302504) antibodies were purchased from Abcam. All antibodies were used following the manufacturer’s instructions.
5
ZIKV Infection Effects on Interferon Response
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Western blot analysis was conducted on total protein extracted from mock and ZIKV-infected SC grown in 6-well tissue culture plates as described previously (Kumar et al., 2013 (link)). β-actin and IRF3 were used as loading controls and were detected using β-actin mouse monoclonal antibody (8H10D10, Cell Signaling Technology; 1:1,000 dilution) and IRF3 rabbit polyclonal antibody (11904, Cell Signaling Technology; 1:1,000 dilution). MX1 was detected using MX1 rabbit polyclonal antibody (sc-50509, Santa Cruz; 1:1,000 dilution). Secondary antibodies (1:10,000 dilution) were conjugated with IRDye 800 and IRDye 680 (Li-Cor Biosciences), and blots were scanned using an Odyssey infrared imager. For IFN-β ELISA, secreted IFN-β in SC supernatant was quantified using an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems DY814-05) according to the manufacturer protocol.
6
Immunoblotting for cell signaling
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The NDR1 (sc-365555) antibody was purchased from Santa Cruz Technology. The phospho-NDR1/2 (Thr444/Thr442) antibody was purchased from Affinity Biosciences. All the following antibodies were purchased from Cell Signaling Technology: E-cadherin (#14472), PARP (#9532), cleaved-PARP (#5625), phospho-Thr (#9386), β-actin mouse monoclonal antibody (#3700), and secondary antibody against mouse and rabbit. Bcl-2 (A19693), Bax (A19684), N-cadherin (A19083), Vimentin (A19607), Survivin (A1551), YAP (A1002), and phospho-YAP-Ser127 (AP0489) were purchased from ABclonal. All antibodies were used following the manufacturer’s instructions.
7
Western Blot Analysis of SPRY4 Expression
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Proteins in the sample lysates from different colon cancer cell lines were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis on 10% Criterion-XT BioRad gels, and the separated proteins were transferred to BioRad nitrocellulose membrane. Membranes were then blocked with 5% bovine serum albumin (BSA) at room temperature for 1 h followed by overnight incubation with primary SPRY4 rabbit polyclonal antibody at 1:500 dilution (Abcam Cat#ab59785) and β-actin mouse monoclonal antibody at 1:1000 (Cell Signalling Cat# 3700S) at 4°C. Next day, membranes were incubated for 1 h at room temperature with appropriate horseradish peroxidase-coupled secondary antibodies. Signals were detected by using the Super signal® West femto maximum sensitivity substrate (Pierce), and images were visualized by BioRad ChemiDoc XRS imager.
8
Oxidative Stress and Apoptosis Assays
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Ethanol, sodium dodecyl sulphate (SDS), MDA, thiobarbituric acid (TBA), N,N,N-,N-Tetramethylethylenediamine (TEMED), and β-mercaptoethanol (β-ME) were provided by Merck. phenylmethanesulfonyl fluoride (PMSF), complete protease inhibitor cocktail and reduced GSH were gained from Sigma-Aldrich. Rabbit monoclonal caspase-3 antibody, rabbit monoclonal caspase-8 antibody, rabbit monoclonal caspase-9 antibody, rabbit monoclonal Bcl-2 antibody, rabbit polyclonal Bax antibody, antirabbit IgG, horseradish peroxidase-conjugated antibody, mouse monoclonal β-actin antibody, anti-mouse IgG, and horseradish peroxidase-conjugated antibody were purchased from cell signaling. Polyvinylidene fluoride (PVDF) membrane and Bradford protein assay kit were obtained from Bio- Rad. BCA protein assay kit and Western blotting detecting reagents (ECL), was supplied by Pierce. TNF-α Rat ELISA Kit, IL-6 Rat ELISA Kit and Express one-step SYBR R Green ER™kit were provided from Invitrogen. Further chemicals were of the highest grade commercially available.
9
Western Blot Analysis of Elk1 Overexpression
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At 48 h post transfection of 5×105 SH-SY5Y cells, total protein was extracted. Equal amounts (15 μg protein/lane) of proteins obtained from whole cell extracts of untransfected and Elk1 transfected samples were separated by SDS-PAGE and transferred onto nitrocellulose membrane by Trans-Blot Turbo Blotting System (Bio-Rad). Membrane was blocked in 5% skim milk powder/TBST (Tris buffered saline (TBS) containing 0.1% Tween 20) for 1 h at room temperature, and then incubated with the following primary antibodies at indicated dilutions overnight at 4°C; rabbit monoclonal His-tag antibody (1:1000, Cell Signaling Technology), mouse monoclonal spastin antibody (1:1000, Sigma), rabbit polyclonal katanin-p60 antibody (1:1000, ATLAS), mouse monoclonal p27 antibody (1:500, Santa Cruz), mouse monoclonal HuR antibody (1:500, Santa Cruz), mouse monoclonal β-Actin antibody (1:1000, Cell Signaling Technology), rabbit monoclonal GAPDH antibody (1:1000, Cell Signaling Technology), rabbit monoclonal β-tubulin antibody (1:1000, Cell Signaling Technology) in 5% skim milk powder/TBST. Membranes were then incubated with Horseradish peroxidase conjugated goat anti-rabbit or anti-mouse IgG secondary antibodies (1:3000, Cell Signaling Technology) for 1 h at room temperature. Bands were visualized using Visualizer Western Blot Detection Kit (Millipore) and ChemiDoc Imaging System (Bio-Rad).
10
Apoptosis and Oxidative Stress Assay
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This study was performed using the following items: Annexin V-FITC apoptosis/necrosis detection kit and fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA) (Abcam, USA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) and propidium iodide (PI) ( Sigma, USA), RPMI 1640, FBS (fetal bovine serum), penicillin and streptomycin (Gibco, USA), rabbit polyclonal Hsp70 antibody, rabbit polyclonal Hsp90 antibody, anti-rabbit IgG horseradish peroxidase-conjugated antibody, mouse monoclonal β-actin antibody, (Cell signaling, USA), polyvinylidene fluoride (PVDF) membrane (Bio-Rad, USA) and enhanced chemiluminescnces (ECL) reagent for western blot detection (Pierce).
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