Antibodies were obtained from the following sources; anti-MAP2, anti-beta-tubulin, and anti-synaptophysin from Cell Signaling Inc. (Beverly, MA), β-tubulin from LI-COR, Odyssey (Lincoln, NE), anti-SRSF1 from Invitrogen, anti-SRSF2 from Sigma Aldrich, anti-MCL1, anti-SRSF3, anti-SRSF4, and anti-HNRNP A1 from Santa Cruz Biotech, and anti-Grb2 from BD Biosciences. Mammalian protease inhibitors were obtained from Sigma-Aldrich (St Louis, MO). Bradford reagent was from BioWorld (Dublin, OH). MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) was obtained from Thermo Fisher Scientific (#M6494).
Anti beta tubulin
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-beta-tubulin is a primary antibody that specifically recognizes the beta-tubulin protein. Beta-tubulin is a key component of microtubules, which are cytoskeletal structures involved in various cellular processes, such as cell division, intracellular transport, and cell motility.
Automatically generated - may contain errors
Lab products found in correlation
Anti egfp, by Cell Signaling Technology (2 mentions)
Anti bcl6, by Santa Cruz Biotechnology (2 mentions)
Anti mouse alexa fluor 633, by Thermo Fisher Scientific (2 mentions)
Anti v5 tag, by Thermo Fisher Scientific (2 mentions)
Anti histone h3, by Cell Signaling Technology (2 mentions)
Anti hdac1, by Cell Signaling Technology (2 mentions)
Anti hsp90, by Cell Signaling Technology (2 mentions)
Anti grb2, by BD (1 mentions)
Anti map2, by Cell Signaling Technology (1 mentions)
Anti srsf3, by Santa Cruz Biotechnology (1 mentions)
Mammalian protease inhibitor, by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions)
Anti srsf1, by Thermo Fisher Scientific (1 mentions)
Anti srsf2, by Merck Group (1 mentions)
Anti mcl 1, by Santa Cruz Biotechnology (1 mentions)
Anti synaptophysin, by Cell Signaling Technology (1 mentions)
β tubulin, by LI COR (1 mentions)
Anti hnrnp a1, by Santa Cruz Biotechnology (1 mentions)
Bradford reagent, by Bioworld Technology (1 mentions)
Cfx96 system, by Bio-Rad (1 mentions)
8 protocols using anti beta tubulin
1
Antibody Sources and Reagents
2
Comprehensive Antibody Panel for Protein Analysis
3
Silencing Pin1 in Hep3B Cells
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Hep3B cells were seeded in 6‐well plates at 2 × 105 cells/well. DP7‐C and GalNAc‐Pin1 siRNA complex was incubated with the Hep3B cells at doses of 1 μg GalNAc‐Pin1 siRNA. Hep3B cells were harvested 48 hours later for analysis. Total RNA was extracted with a kit (ForeGene). qPCR reactions were performed on a Bio‐Rad CFX96 system.
The western blot results better showed the silencing efficiency of the siRNA. The protein samples were probed with anti‐GAPDH (Cell Signaling Technology, CST), anti‐Pin1 (CST), anti‐Beta Tubulin (CST), anti‐Lamin B (CST), and anti‐Exportin 5 (CST) antibodies. The cells were incubated with primary antibodies at 4°C overnight. Then, the membranes were incubated with secondary antibodies (CST) for 60 minutes and developed using the ECL detection system (Vazyme).
The western blot results better showed the silencing efficiency of the siRNA. The protein samples were probed with anti‐GAPDH (Cell Signaling Technology, CST), anti‐Pin1 (CST), anti‐Beta Tubulin (CST), anti‐Lamin B (CST), and anti‐Exportin 5 (CST) antibodies. The cells were incubated with primary antibodies at 4°C overnight. Then, the membranes were incubated with secondary antibodies (CST) for 60 minutes and developed using the ECL detection system (Vazyme).
4
Comprehensive Antibody Panel for Protein Analysis
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The following antibodies were used in this study: anti-BCL6 (Santa Cruz Biotechnology, sc-7388), anti-beta-tubulin (Cell Signaling, 2146S), anti-Hsp90 (Cell Signaling, 4874S), anti-HDAC1 (Cell Signaling, 2062S), anti-Histone H3 (Cell Signaling, 12648S), anti-eGFP (Cell Signaling, 2956), anti-V5-tag (ThermoFisher Scientific, MA5–15253), anti-Streptavidin (Sigma, 71591–3), anti-Mouse 800CW (LI-COR Biosciences, 926–32211), anti-Rabbit 680LT (LI-COR Biosciences, 925–68021), anti-mouse Alexa Fluor 633 (ThermoFisher Scientific, A-21052), and Alexa anti-mouse 488 (Biolegend, 405319).
5
TTC-Based Apoptosis Analysis Workflow
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HE (Nanjing Spring & Autumn Biological Engineering Co. Ltd) with a purity greater than 98% was dissolved in 2% dimethyl sulfoxide (DMSO) to generate stock solutions. 2,3,5- triphenyltetrazolium chloride (TTC) was purchased from Sigma-Aldrich; rabbit beta-actin polyclonal antibody, rabbit anti-Bcl-2, and rabbit anti-Bax were purchased from Bioworld Technology Inc. (St. Louis Park, MN, USA). Rabbit beta-IKK polyclonal antibody were purchased from Cell Signalling Technology; RIPA lysis buffer was purchased from Millipore (Billerica, MA, USA). A BCA Protein Assay Kit was purchased from Thermo-Fisher Scientific (MA, USA); anti-beta-tubulin, anti-NF-κB (p-p65), anti-SAPK/JNK, anti-phospho-SAPK/JNK, and goat anti-rabbit secondary antibody were purchased from Cell Signaling Technology. Goat anti-Iba-1 was purchased from Abcam (1:500); Trizol was purchased from Invitrogen (USA). A PrimeScript RT reagent kit was purchased from Takara (Dalian, China); Lv-MLK3 and Lv-con were purchased from GeneChem (Shanghai China). A SYBR green Kit was purchased from Applied Biosystems (Foster City, CA, USA) and the ECL chemiluminescence system was purchased from Thermo Company (Rockford, IL, USA).
6
Quantification of Insulin Signaling Proteins
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For each genotype 100 adult heads were homogenized in RIPA I buffer [50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, protease inhibitor cocktail (Roche Diagnostic; Basel, Switzerland)] on ice. After centrifugation, loading buffer was added to the supernatant and the probes were loaded on a SDS-PAGE followed by Western blotting. The extracts were probed with guinea pig anti-CycG (1:400) [46 (link)]. As loading controls we used anti-beta-Tubulin (1:50) (E7 DSHB; developed by M. Klymkowsky) and anti-Erk1/2 antibodies (1:1000) (Cell Signaling Technology; Danvers MA, USA), as tubulin levels might change when InR signaling is influenced [15 (link)]. The amount of total and phosphorylated protein was determined with rabbit anti-4E-BP (1:100, gift from G. Tettweiler) [58 (link)], rabbit anti-Akt1 (1:250), rabbit anti-p70 S6 kinase (1:100), rabbit anti-Phospho-Akt (1:250), rabbit anti-Phospho-p70 S6 kinase (1:100) and rabbit anti-Phospho-4E-BP (1:100) (all from Cell Signaling Technology; Danvers MA, USA).
7
Western Blot Protocol for Dicer and Ago2
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For western blotting, we used the protocol previously described in Mori et al., 2008 [22] . The antibodies were: anti-Dicer (13502, Abcam®), anti-Ago2 (2897, Cell Signaling®), anti-beta-tubulin (2146, Cell Signaling®), anti-rabbit IgG-HRP (RPN4301, GE®). Scion Image® software was used for the quantification of bands by densitometry. We used beta-tubulin or Ponceau S dye (Sigma–Aldrich®) as loading controls.
8
Comprehensive Antibody Characterization Protocol
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The polyclonal chicken-derived anti-Gal-3 antibody was produced in our laboratory (Toledo et al. 2014) (link). The human recombinant Gal-3 (hr-Gal-3) protein was prepared as previously described (Arthur, Rodrigues, et al. 2015) . Antibodies anti-dystrophin (sc-153760), antidesmin (sc-7559), anti-myogenin (sc-576) and anti-pCREBSer133 (sc-9198) were purchased from Santa Cruz Biotechnology (Santa Cruz, Dallas, TX, USA); anti-pAktSer473 (#9271) and anti-betatubulin (#2146) were purchased from Cell signaling (Danvers, MA, USA); anti-beta1-Integrin (CSAT) and anti-OPN were purchased from DSHB (Developmental Studies Hybridoma Bank at University of Iowa, IA, USA); anti-MyoD (clone 5.8A, ThermoFisher, Waltham, MA, USA); anti-Pax7 (MAB020 2F-12H4, Millipore, Burlington, MA, USA); donkey anti-rabbit-Alexa_546, donkey anti-mouse-Alexa_546, Goat anti-Chicken-Alexa_488 from Molecular Probes ® -Thermo Fisher Scientific; donkey immunoglobulin G (IgG), donkey anti-goat horseradish peroxidase (HRP), donkey anti-mouse HRP, donkey anti-rabbit HRP and Donkey anti-chicken HRP were purchased from Jackson ImmunoResearch (West Groove, PA, USA). DAPI (4,6-Diamidino-2-Phenylindole, Dihydrochloride) and Hoechst 33342, Trihydrochloride, Trihydrate from Thermo Scientific. The negative control was added in Supplementary Figure S1C, a-f).
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