After lysis with lysis buffer, 20 μg of protein per sample was loaded onto a lane of a sodium dodecyl sulphate-polyacrylamide gel for electrophoresis. The proteins were electrotransferred to polyvinylidene fluoride membranes. The transferred membrane was treated with blocking buffer and incubated with primary antibodies (E-cadherin [20874-1-AP; Proteintech, Chicago, IL, USA)], N-cadherin [22018-1-AP; Proteintech, Chicago, IL, USA], Cyclin D1 [60186-1-lg; Proteintech, Chicago, IL, USA], β actin [MAB1501R; Millipore, Burlington, VT, USA]) for 2 h at room temperature. Secondary antibodies (goat anti-rabbit [AP132P; Millipore, Burlington, VT, USA] and goat anti-mouse [AP124P; Millipore, Burlington, VT, USA]) were then added, and the cells were incubated for 90 min at room temperature. An enhanced chemiluminescence solution (205–14,621; Revvity, Burlington, VT, USA) was used to detect specific bands using a MINICHEMI (Thermo) system.
β actin mab1501r
Manufactured by Merck Group
Sourced in United States
β-actin (MAB1501R) is a monoclonal antibody that recognizes the β-actin isoform, a highly conserved cytoskeletal protein found in all eukaryotic cells. This antibody can be used in various applications to detect and study the expression and distribution of β-actin in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Minichemi, by Thermo Fisher Scientific (1 mentions)
N cadherin 22018 1 ap, by Proteintech (1 mentions)
E cadherin 20874 1 ap, by Proteintech (1 mentions)
Cyclind1 sc 8396, by Santa Cruz Biotechnology (1 mentions)
Dkk 1 sc 25516, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 488 conjugated antibody, by Thermo Fisher Scientific (1 mentions)
Ki67 gtx16667, by GeneTex (1 mentions)
Erk2 sc 154, by Santa Cruz Biotechnology (1 mentions)
Alkaline phosphatase conjugated antibodies, by Promega (1 mentions)
Dusp6 ab76310, by Abcam (1 mentions)
H3k9me3, by Active Motif (1 mentions)
H3k9ac, by Active Motif (1 mentions)
Gapdh sc 25778, by Santa Cruz Biotechnology (1 mentions)
Image lab software, by Bio-Rad (1 mentions)
Chemidoc imaging system, by Bio-Rad (1 mentions)
Chemidoc touch imaging system, by Bio-Rad (1 mentions)
Hif 1α, by BD (1 mentions)
4 protocols using β actin mab1501r
1
Western Blot Analysis of Cell Signaling Proteins
2
Immunoblotting Analysis of Signaling Proteins
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Radioimmunoprecipitation (RIPA) buffer with phosphatase-inhibitor cocktails and protease-inhibitor was used to obtain cell lysates. Immunoblotting was carried out as previously described [26 (link)]. The following antibodies were used: β-catenin (F5682) from BD Biosciences (Franklin Lakes, NJ, USA); CyclinD1 (SC-8396) and DKK-1 (SC-25516) from Santa Cruz Biotechnology (Dallas, TX, USA); GSK-3β (9315S) and p-GSK-3β (9336) from Cell Signaling Technology (Cell Signaling Technology, Beverly, MA, USA); β-actin (MAB1501R) from Millipore (Billerica, MA, USA); α-tubulin (GTX628802) and SP1 (GTX110593) from GeneTex (Irvine, CA, USA).
3
Immunoblotting and Immunofluorescence Protocols
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Primary antibodies were obtained from the following sources: DUSP6 (ab76310) from Abcam (Toronto, Canada), β‐actin (MAB1501R) from Millipore (Oakville, Canada), ERK2 (sc‐154) and MEK2 (sc‐524) from Santa Cruz Biotechnology (Santa Cruz, CA), phosphorylated ERK1/2 (M8159) from Sigma‐Aldrich (Oakville, Canada), phosphorylated MEK1/2 (9121) from Cell Signaling Technology (Danvers, MA, USA), chromogranin A (20085) from ImmunoStar (Hudson, WI), proliferating cell nuclear antigen (PCNA; 18197) from Abcam and Ki67 (GTX16667) from Genetex (Irvine, CA). Horseradish peroxidase antibodies were obtained from GE Healthcare Life Sciences (Mississauga, Canada) and alkaline phosphatase‐conjugated antibodies from Promega (Madison, WI). For immunofluorescence, Alexa Fluor 488 conjugated antibodies were obtained from Molecular Probes (Waltham, MA). Other materials were purchased from Sigma‐Aldrich unless stated otherwise.
4
Comprehensive Western Blotting Methodology
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Western blotting was conducted as described previously3 (link),6 (link). Immunoblots were visualized using the ChemiDoc Imaging System (Bio-Rad). After chemiluminescent reaction, blots were visualized with a ChemiDoc Touch Imaging System (Bio-Rad). The signal intensities of the captured images were analyzed with Image Lab Software (Bio-Rad, version 5.2.1). Results of Western analyses shown are representatives of at least three independent experiments. Antibodies used were OXPHOS Cocktail (MS501) from MitoSciences (1:1000); β-actin (MAB1501R) from Millipore (1:5000); GAPDH (sc-25778) from Santa Cruz (1:5000); H3 (39763; 1:10,000), H3K9me3 (61013; 1:1000), and H3K9Ac (61251; 1:1000) from Active Motif; eIF2α (2103S), p-eIF2α (9721S), p70S6K (9202), phospho-p70S6K (Thr389, 9234) and ATF4 (11815S) from Cell Signaling (1:1000 for all); Pan Acetyl-H3 (ab47915; 1:1000) from Abcam; HIF-1α (610958; 1:3000) from BD Biosciences; and ASNS (14681-1-AP; 1:3000) from Proteintech. The anti-ASS1 antibody was a gift from L.-J. Shen6 (link).
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