Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 °C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubated with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed using an Odyssey imaging system (Li-Cor, Lincoln, NE).
Ab133695
Manufactured by Abcam
Sourced in United Kingdom
Ab133695 is a protein that functions as a structural component of the microtubule cytoskeleton. It is commonly used in research applications involving the study of cellular architecture and dynamics.
Automatically generated - may contain errors
Lab products found in correlation
Bca assay kit, by Thermo Fisher Scientific (5 mentions)
Ab129087, by Abcam (4 mentions)
Ab18950, by Abcam (4 mentions)
Ab184955, by Abcam (4 mentions)
Ab191563, by Abcam (4 mentions)
Ab6721, by Abcam (4 mentions)
Odyssey imaging system, by LI COR (4 mentions)
Odyssey blocking reagent, by LI COR (4 mentions)
Pvdf membrane, by Merck Group (4 mentions)
Ab213480, by Abcam (1 mentions)
Ab49900, by Abcam (1 mentions)
Anti phospho iκbα ser32 36 5a5, by Cell Signaling Technology (1 mentions)
Goat anti rabbit igg h l hrp conjugate, by Abcam (1 mentions)
Anti phospho sapk jnk thr183 tyr185 81e11, by Cell Signaling Technology (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Ab32072, by Abcam (1 mentions)
Ab205718, by Abcam (1 mentions)
Gtx127375, by GeneTex (1 mentions)
6 protocols using ab133695
1
Western Blot Analysis of Synaptic Proteins
2
Immunoblot analysis of signaling proteins
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Cells were lyzed with lysis buffer (20 mM Tris-HCl [pH 7.5], 150 mM NaCl, 1 mM EDTA, 1% NP40) containing Complete Protease Inhibitor Cocktail (Roche). The cell lysates were separated by standard SDS-PAGE (e-PAGEL, ATTO, Tokyo, Japan) and analyzed by immunoblotting. The following antibodies were used for immunoblotting: anti-PD-L1 (ab213480, Abcam, Cambridge, UK), anti-EGR1 (ab133695, Abcam), anti-c-Myc antibody (Y69) (ab32072, Abcam), goat anti-rabbit IgG (H+L)-HRP Conjugate (Bio-Rad), anti-phospho-p38 antibody (Thr 180/Tyr 182; sc-17852-R, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-phospho-SAPK/JNK (Thr183/Tyr185; 81E11, Cell Signaling Technology, Danvers, MA, USA), anti-IκBα mouse antibody (L35A5, Cell Signaling Technology), anti-phospho-IκBα (Ser32/36; 5A5, Cell Signaling Technology), anti-β-actin (C4) (sc-47778, Santa Cruz Biotechnology), and anti-β-actin-HRP (AC-15) (ab49900, Abcam). The Western HRP Substrate (Pierce ECL Western Blotting Substrate, Thermo Fisher Scientific) was used for the development of positive signals, and chemiluminescence was detected using a ChemiDoc XRS+ System (Bio-Rad, Hercules, CA, USA).
3
Western Blot Analysis of Autophagy Markers
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Cell lysate derived from Hep3B or Hep3B/So was prepared using lysis buffer (PBS + 1% NP40 + 0.1% SDS + 5 mmol/L EDTA + 0.5% sodium deoxycholate + 1 mmol/L sodium orthovanadate), and protein concentration was determined by BCA Assay Kit (ThermoScientific). 40 μg protein samples were separated on 10% SDS‐PAGE and then electrotransferred onto PVDF membrane, which was blocked by 5% skimmed milk at room temperature for 1 hour Next, the membrane was incubated with the primary antibodies LC3I (GTX17380, 1:1000; GeneTex) and LC3II (GTX127375, 1:3000; GeneTex), EGR1 (ab133695, 1:1000; Abcam) at 4°C overnight. The membrane was rinsed in TBST solution and then incubated with the secondary goat anti‐rabbit antibody (ab205718, 1:5000; Abcam) for 2 hours. The signal was determined using enhanced ECL (Perkin Elmer) by ImageQuant LAS 4000 (ImageQuantTL, Imagemoster 2D Platinum 6.0; GE Healthcare Life Science).
4
Western Blot Analysis of Neuronal Proteins
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Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 o C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubated with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed using an Odyssey imaging system (Li-Cor, Lincoln, NE).
5
Western Blot Analysis of Neural Proteins
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Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 o C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubation with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed by using an Odyssey imaging system (Li-Cor, Lincoln, NE).
6
Western Blot Analysis of Neuronal Proteins
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+ Expand
Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 o C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubated with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed using an Odyssey imaging system (Li-Cor, Lincoln, NE).
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