Whole cell extracts (30 μL each) were electrophoresed and protein samples in the gels were transferred to a nitrocellulose membrane as previously shown [16 (link)]. Immunoreactive bands of HDAC2, HDAC5 and β-actin were detected with anti-HDAC2 antibody (Santa Cruz Biotechnology, Santa Cruz, CA), anti-HDAC5 antibody (Santa Cruz Biotechnology) and anti-β actin antibody (Abcam, Cambridge, UK), respectively. Band densities of HDAC2 and HDAC5 in each sample were normalized with that of the β-actin or glyceraldehyde 3-phosphate dehydrogenase (GAPDH).
Anti hdac5 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United Kingdom
The Anti-HDAC5 antibody is a laboratory reagent used to detect and study the HDAC5 protein. HDAC5 is a class II histone deacetylase enzyme that plays a role in regulating gene expression. The antibody can be used in various applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to identify and quantify the HDAC5 protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Eco real time pcr system, by Illumina (2 mentions)
Histone3 antibody, by Abcam (2 mentions)
Domitol, by Meiji Seika Pharma (2 mentions)
Vetorphale, by Meiji Seika Pharma (2 mentions)
Dormicum, by Astellas Pharma (2 mentions)
Anti hdac2 antibody, by Santa Cruz Biotechnology (1 mentions)
Anti β actin antibody, by Abcam (1 mentions)
3 protocols using anti hdac5 antibody
1
Western Blot Analysis of HDAC2 and HDAC5
2
CGRP Enhances Npas4 Transcription in Mice
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A total of 50 mice were used in this experiment. CGRP or saline was administered i.c.v. for 24 h before cross-linking. The mice were deeply anesthetized with three types of mixed anesthetic agents52 (link), medetomidine hydrochloride (Domitol, Meiji Seika Pharma Co., Ltd., Tokyo, Japan, 0.3 mg/kg), midazolam (Dormicum, Astellas Pharma Inc., Tokyo, Japan, 4.0 mg/kg), and butorphanol (Vetorphale, Meiji Seika Pharma Co., Ltd., 5.0 mg/kg), administered intraperitoneally and perfused transcardially with saline, followed by 1% paraformaldehyde, pH 7.4. The hippocampus was post-fixed for 10 min in 1% paraformaldehyde, and then glycine (330 mM) was added. The chromatin was then sheared into approximately 0.5–1-kb fragments and immunoprecipitated using an anti-HDAC5 antibody (1:100 for the hippocampal sample, Santa Cruz Biotechnology), acetylated Histone3 antibody (1:100, Abcam plc) or Histone3 antibody (1:100, Abcam plc). Immunoprecipitated DNA fragments were amplified and quantitated by qPCR (Eco Real-Time PCR System (Illumina Inc.), using PCR primers specific for Npas4, which were designed around the putative enhancer regions of Npas4 (Table 2 ).
Oligonucleotide sequences for ChIP.
| Forward | Reverse | |
|---|---|---|
| Npas4 | 5ʹ-TGCCAGGCTATTTTTGGTTC-3ʹ | 5ʹ-CTGTATGCCCCCAATGTCTC-3ʹ |
3
Epigenetic Regulation of Npas4 Expression
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CGRP or saline was administered i.c.v. for 24 hr before cross-linking. The mice were deeply anesthetized with three types of mixed anesthetic agents (49) , medetomidine hydrochloride (Domitol, Meiji Seika Pharma Co., Ltd., Tokyo, Japan, 0.3 mg/kg), midazolam (Dormicum, Astellas Pharma Inc., Tokyo, Japan, 4.0 mg/kg), and butorphanol (Vetorphale, Meiji Seika Pharma Co., Ltd., 5.0 mg/kg), administered intraperitoneally and perfused transcardially with saline, followed by 1% paraformaldehyde, pH 7.4. The hippocampus was post-fixed for 10 min in 1% paraformaldehyde, and then glycine (330 mM) was added. The chromatin was then sheared into approximately 0.5-1-kb fragments and immunoprecipitated using an anti-HDAC5 antibody (1:100 for the hippocampal sample, Santa Cruz Biotechnology), acetylated Histone3 antibody (1:100, Abcam plc) or Histone3 antibody (1:100, Abcam plc). Immunoprecipitated DNA fragments were amplified and quantitated by qPCR (Eco Real-Time PCR System (Illumina Inc.), using PCR primers specific for Npas4, which were designed around the putative enhancer regions of Npas4 (Table 2).
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