Antibodies (Abs) used in immunoblotting included anti-phospho-BAD (S112), anti-BAD, anti-phospho-p70S6K (T389), anti-p70S6K, anti-4E-BP1 (T37/46), anti-4E-BP1, and anti-c-Myc, and were purchased from Cell Signaling Technology (Danvers, MA). Anti-TRAF3 (H122) Ab was from Santa Cruz Biotechnology (Dallas, TX) and the anti-mouse actin Ab was from Sigma (St. Louis, MO). The anti-Pim2 and anti-β-actin Abs were from Thermo Fisher (Waltham, MA). Rabbit anti-K48 polyubiquitin Ab was purchased from Abcam (Cambridge, MA). HRP-conjugated goat-anti-mouse IgG, and goat-anti-rabbit Ig Abs were from Jackson ImmunoResearch Laboratories (West Grove, PA). Cucurbatacin I was purchased from Santa Cruz Biotechnology. SGI-1776 was graciously provided by Dr. David Bearss (University of Utah, Salt Lake City, UT) or purchased from ApexBio (Houston, TX). TP-3654 was also a gift from Dr. Bearss. 10058-F4 was from Tocris (Bristol, UK). JQ1 was purchased from Cayman Chemical (Ann Arbor, MI).
Anti mouse actin ab
Manufactured by Merck Group
Anti-mouse actin Ab is a primary antibody that specifically binds to actin, a cytoskeletal protein found in mouse cells. It can be used in various laboratory techniques to detect and study the distribution and expression of actin.
Automatically generated - may contain errors
Lab products found in correlation
Anti 4e bp1, by Cell Signaling Technology (1 mentions)
Anti c myc, by Cell Signaling Technology (1 mentions)
Hrp conjugated goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Immobilon p polyvinylidene fluoride, by Merck Group (1 mentions)
Image gauge software, by Fujifilm (1 mentions)
Horseradish peroxidase conjugated donkey anti mouse and anti rabbit, by GE Healthcare (1 mentions)
Universal hood 2, by Bio-Rad (1 mentions)
Recombinant murine m csf, by R&D Systems (1 mentions)
Pgex 4t 1, by GE Healthcare (1 mentions)
Rabbit anti integrin β3, by Cell Signaling Technology (1 mentions)
Mouse anti flag ab, by Merck Group (1 mentions)
4 protocols using anti mouse actin ab
1
Western Blot Analysis of Cell Signaling
2
Quantifying Pim1 Kinase Isoforms in Stimulated Cells
3
Quantification of BACE1 in Human Brain
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Human brain tissue sections, obtained as indicated in the subsection 2.1., were lysed with 50 μL RIPA buffer: 150 mM sodium chloride, 1% Triton X-100 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris-HCl, 1 mM dithiothreitol, 1 mM sodium orthovanadate, and protease inhibitor cocktail (Roche), pH 8. Membranes were blocked for 1 h at RT with Tween 20-Tris buffer solution (TTBS; 100 mM Tris-HCl, 150 mM NaCl, pH 7.5) plus 5% skimmed milk. Then, membranes were incubated overnight (o.n.) at 4°C with 1 : 4,000 mouse anti-actin Ab (Sigma) or 1 : 1,000 rabbit anti-BACE1 Ab. 1 : 2,000 secondary Abs were horseradish peroxidase-conjugated donkey anti-mouse and anti-rabbit (GE Healthcare), which were incubated for 1 h at RT. Bands were visualized with Super Signal (Pierce) and analyzed with the Quantity One system in a BioRad Universal Hood II.
4
Recombinant murine M-CSF and GST-RANKL production
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Recombinant murine M-CSF was obtained from R&D Systems. Glutathione-S-transferase (GST)-RANKL was expressed in our laboratory. The GST-RANKL vector was constructed by PCR cloning a SalI/NotI fragment corresponding to amino acids 158–316 of murine RANKL from osteoblast cDNA (forward primer, 5′-ATGTGGCCCGTCGACGCAAGCCTGAGGCCCAGCC-3′; reverse primer, 5′-AAACATCTAGGGCGGCCGCGCTAATG-3′) and cloning into pGEX-4T-1 (GE Healthcare). Both strands were sequenced. High-expressing BL21 (Stratagene) cells were lysed under nondenaturing conditions and GST-RANKL was purified over a glutathione-Sepharose column followed by ion exchange chromatography (Lam et al., 2000 (link)). The sources of antibodies were as follows: rabbit anti-integrin β3 (Cell Signaling Technology); mouse antiphosphotyrosine (4G10), rabbit anti-FcRγ from Millipore; mouse anti-NFATc1 and rabbit anti-Syk (N19; Santa Cruz Biotechnology, Inc.); mouse anti-Flag Ab, rabbit anti-Flag Ab, mouse anti-Talin Ab, and mouse anti-actin Ab (Sigma-Aldrich); mouse anti-HA Ab (Covance); goat anti-OSCAR Ab (R&D Systems); rabbit anti-β1 Ab (Abcam). All other chemicals were obtained from Sigma-Aldrich.
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