The following antibodies were used: HDAC5, HDAC7, and MyD88 (Abcam, Cambridge, MA); active caspase-3, p-P70 S6K (T389), and P70 S6K antibodies (Cell Signaling, Danvers, MA); α-tubulin and FLAG antibodies (Sigma-Aldrich, St. Louis, MO). Polyclonal anti-HDAC4 antibody was described previously (Zhao et al., 2001 (link)). GSK3 inhibitor SB216763, iNOS inhibitor NAME, Griess reagent, and LPS were purchased from Sigma-Aldrich.
Hdac7
Manufactured by Abcam
Sourced in United States, United Kingdom
HDAC7 is a human histone deacetylase enzyme that is involved in the regulation of gene expression. It catalyzes the removal of acetyl groups from lysine residues on histone proteins, leading to the condensation of chromatin and transcriptional repression.
Automatically generated - may contain errors
Lab products found in correlation
Hdac4, by Cell Signaling Technology (3 mentions)
Supersignal west pico, by Thermo Fisher Scientific (2 mentions)
Femto ecl, by Thermo Fisher Scientific (2 mentions)
Hdac3, by Cell Signaling Technology (2 mentions)
Hdac5, by Cell Signaling Technology (2 mentions)
Cd166, by Abcam (2 mentions)
Cd44 2c5, by R&D Systems (2 mentions)
Hdac6, by Novus Biologicals (2 mentions)
Hdac8, by Novus Biologicals (2 mentions)
Hdac10, by Novus Biologicals (2 mentions)
Gradient sds page gel, by Bio-Rad (2 mentions)
Hdac9, by Abcam (2 mentions)
Hdac1, by Cell Signaling Technology (2 mentions)
Hdac2, by Cell Signaling Technology (2 mentions)
Sirt1, by Abcam (2 mentions)
β actin, by Abcam (2 mentions)
Ach3k27, by Abcam (2 mentions)
Ph3s10, by Abcam (2 mentions)
Ac h3k9, by Abcam (2 mentions)
Lysis buffer, by Keygen Biotech (2 mentions)
8 protocols using hdac7
1
Investigating HDAC and Apoptosis Signaling
2
Comprehensive Protein Expression Profiling
3
Protein Expression Analysis of Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-cell lysates, harvested with RIPA buffer containing protease and phosphatase inhibitors, were run on 4–15 % SDS-PAGE gradient gels (Biorad, Hercules, CA, USA). Blots were blocked in 5 % milk, then probed with antibodies; CD326 (Cell Signaling, Danvers, MA, USA; #2929 S), CD166 (Abcam#ab49496), CD44(2c5) (R&D#BBA10), BMI-1 (Cell Signaling#2830 S), HDAC1 (Cell Signaling#5356), HDAC2 (Cell Signaling#5113), HDAC3 (Cell Signaling#3949), HDAC4 (Cell Signaling#7628), HDAC5 (Cell Signaling#2082), HDAC6 (Novus, Littleton, CO, USA; #91805), HDAC7 (Abcam#ab12174), HDAC8 (Novus#91807), HDAC9 (Abcam#ab18970), HDAC10 (Novus#91801). Blots were developed using SuperSignal West Pico or Femto ECL (ThermoScientific, Waltham, MA, USA). All experiments were repeated a minimum of three times.
4
Protein Detection Across Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies against the following proteins were used for our studies: HDAC7 (#ab53101) and AKAP12 (#ab49849) from Abcam (Cambridge, UK); acetylated lysine (#9441), STAT3 (#8768), Phospho-STAT3 (Tyr705)(#9145), Acetyl-Stat3 (Lys685)(#2523), JAK1(#3344), JAK2(#3230), PKC(#2056), p53(#2524) from Cell Signaling (Danvers, MA, USA); anti-HA(#H3663), anti-FLAG(#F1804) and anti-β-Actin(A3854) from Sigma-Aldrich (St Louis, MI, USA); Myc (#SC-40) from Santa Cruz (Santa Cruz, CA, USA).
Standard Western blot protocol was adopted. Images were acquired with Tanon-5200 and the density of bands was determined with Image J. The methods for immunoprecipitation was previously described [31 (link)].
Standard Western blot protocol was adopted. Images were acquired with Tanon-5200 and the density of bands was determined with Image J. The methods for immunoprecipitation was previously described [31 (link)].
5
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was extracted from intestinal tissue samples using lysis buffer (KeyGEN, Nanjing, China). The protein concentrations of each sample were calculated with the BCA protein assay kit. Protein was used for western blot analysis, after adding 6× concentrated sample buffer (0.5M Tris, 30% glycerol, 10% SDS, 0.6M DTT, 0.012% bromophenol blue) and heating the samples for 5 min at 95°C. Proteins in supernatants were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene fluoride membranes (PVDF) membranes. Next, the membranes were incubated with the primary antibodies for β-actin (Abcam, MA, United States), SIRT1, HDAC7, acH3, acH3K9, acH3K27, and pH3S10 (Abcam, MA, United States) overnight at 4°C after blocked with Tris-buffered saline containing 0.1% Tween-20 (TBS-T) and 5% low-fat milk blocked for 1 h at RT. After washing for 1 h in TBS-T five times, the membranes were incubated for 1 h at RT with horseradish peroxidase (HRP) secondary antibodies. Protein immunoreactive bands were photographed. Each special banding gray value was digitized using ImageJ software, and the gray value of target protein was divided by internal reference β-actin.
6
Protein Expression and Modification Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed with 0.5% NP40 lysis buffer and proteins were blotted following standard protocol. Signals were probed using the chemiluminescence ECL plus reagent (Thermo, Grand Island, NY, USA) and detected using a Typhoon FLA9500 scanner (GE, Fairfield, CT, USA). Primary antibodies were as follows: HDAC1 (abcam, Cambridge, UK), HDAC2 (abcam), HDAC3 (abcam), HDAC4 (CST, Danvers, MA, USA), HDAC5 (Santa Cruz), HDAC6 (CST), HDAC7 (abcam), cleaved caspase-3 (Antibody Revolution, San Diego, CA, USA), cleaved PARP (CST), β-actin (Sigma), acetyl-histone H3 (Millipore), acetylated α-tubulin (abcam), α-tubulin (CST), phospho-p70 S6 kinase (CST) and p70 S6 kinase (CST). LC3, P53, PUMA, HA, and anti-histone H3 antibodies were kindly provided by the Zhao lab of Fudan University (Shanghai, China).
7
TLR4 Silencing via siRNA Lentivirus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A specific small interfering RNA (siRNA) lentivirus vector targeting TLR4 (TLR4-siRNA-lentivirus) was obtained from Shanghai GeneChem (Shanghai, China). The construction and verification of TLR4-siRNA-lentivirus has been described previously. 12 HDAC inhibitor TSA and purified LPS (derived from O55:B5 E. coli) were obtained from Sigma (St Louis, MO, USA). Antibodies against TLR4, HDAC-4, histone H3, acetylated histone H3, histone H4, acetylated histone H4, and GAPDH were purchased from Cell Signaling Technology (Boston, MA, USA). Anti-HDAC-5, HDAC-7, and Thy-1 antibodies were obtained from Abcam (Cambridge, UK). Goat anti-mouse IgG and goat anti-rabbit IgG secondary antibodies were from Santa Cruz Biotechnologies (Santa Cruz, CA, USA).
8
Western Blot Analysis of Protein Expressions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was extracted from intestinal tissue samples using lysis buffer (KeyGEN, Nanjing, China). The protein concentrations of each sample was calculated with the BCA protein assay kit. Protein was used for western blot analysis, after adding 6× concentrated sample buffer (0.5M Tris, 30% glycerol, 10% SDS, 0.6M DTT, 0.012% bromophenol blue) and heating the samples for 5 min at 95°C. Proteins in supernatants were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene uoride membranes (PVDF) membranes. Next, the membranes were incubated with the primary antibodies for β-actin (Abcam, MA, USA), SIRT1, HDAC7, acH3, acH3K9, acH3K27 and pH3S10 (Abcam) overnight at 4°C after blocked with Tris-buffered saline containing 0.1% Tween-20 (TBS-T) and 5% low-fat milk blocked for 1 h at RT. After washing for 1 h in TBS-T ve times, the membranes were incubated for 1 h at RT with horseradish peroxidase (HRP) secondary antibodies.
Protein immunoreactive bands were photographed. Each special banding gray value was digitized using ImageJ software, and the gray value of target protein was divided by internal reference β-actin.
Protein immunoreactive bands were photographed. Each special banding gray value was digitized using ImageJ software, and the gray value of target protein was divided by internal reference β-actin.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!