Hdac6

Manufactured by ABclonal

Sourced in China

HDAC6 is a histone deacetylase enzyme that catalyzes the removal of acetyl groups from lysine residues on histone proteins, which can lead to changes in chromatin structure and gene expression. The core function of HDAC6 is to deacetylate various cellular proteins, including histones, to regulate their activity and cellular processes.

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Lab products found in correlation

Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions) P akt, by Santa Cruz Biotechnology (1 mentions) P ampk, by Santa Cruz Biotechnology (1 mentions) Brain derived neurotrophic factor (bdnf), by Santa Cruz Biotechnology (1 mentions) β actin, by ABclonal (1 mentions) Nuclear cytosol extraction kit, by Applygen (1 mentions) Hoechst 33342 solution, by Dojindo Laboratories (1 mentions) Hydrogen peroxide solution, by Sinopharm (1 mentions) Bovine serum albumin (bsa), by Sangon (1 mentions) Rotary microtome, by Leica (1 mentions) Bx53 microscope, by Olympus (1 mentions) Nlrp3, by Affinity Biosciences (1 mentions)

3 protocols using hdac6

Brain Protein Expression Analysis in Rats

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Rats were killed, and the whole brains were removed. The cortex and hippocampus were collected, and total protein was extracted with RIPA lysis buffer. Nuclear proteins and cytosolic proteins were extracted with a Nuclear-Cytosol Extraction Kit (P1200, Applygen Technologies Inc, Beijing, China). The concentration of total protein was measured by BCA assay. SDS-PAGE gels (10% or 12%) was used for gel electrophoresis according to the molecular weight of the target protein, and the proteins were transferred to 0.45-μm PVDF membrane by wet electroblotting. The protein bands were visualized and captured using an ImageQuant LAS 4000 mini, and the integrated densities were analyzed by ImageJ.
The following antibodies were used: p-AMPK (sc-101630, Santa Cruz), AMPK (sc-19138, Santa Cruz), p-AKT (sc-33437, Santa Cruz), AKT (sc-8312, Santa Cruz), BDNF (sc-546, Santa Cruz), HDAC6 (A11259, Abclonal), β-actin (AC037, Abclonal), and PCNA (sc-7907, Santa Cruz).

Jian W.X., Zhang Z., Zhan J.H., Chu S.F., Peng Y., Zhao M., Wang Q, & Chen N.H. (2020). Donepezil attenuates vascular dementia in rats through increasing BDNF induced by reducing HDAC6 nuclear translocation. Acta Pharmacologica Sinica, 41(5), 588-598.

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HDAC6 Immunofluorescence Staining Protocol

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After dewaxing and rehydration, tissue sections were incubated for antigen retrieval for 10 min and with Triton X-100 for 10 min. The sections were blocked with 5% BSA, incubated in a humidified chamber for 1 h at room temperature, and incubated with antibodies against target proteins in a humidified chamber overnight at 4 °C. The slides were rinsed three times with PBST and incubated with immunofluorescence secondary antibodies and Hoechst in the dark at room temperature for 2 h. The slides were rinsed three times with PBST, mounted with glycerin, and coverslipped. The tissue sections were visualized and captured with a confocal microscope.
The following antibodies were used: HDAC6 (A11259, Abclonal), Alexa Fluor^TM 488-conjugated donkey anti-rabbit IgG (H + L) (A21206, Invitrogen), and Hoechst 33342 solution (H342, Dojindo).

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Histopathological Evaluation of Acute Lung Injury

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The fixed lung samples were embedded in paraffin as previously described [26] (link), and sectioned serially at 5 μm slices by using a rotary microtome (Leica, Nussloch, Germany). Subsequently, the samples were stained with hematoxylin and eosin (H&E) [27, (link)28] (link). Finally, the stained sections were observed and photographed under a BX53 microscope (Olympus, Tokyo, Japan). According to previous studies [29] (link), the scoring system was used to estimate the severity of acute lung injury as shown in Table 1.
Immunohistochemistry. For immunohistochemistry, the lung tissue sections were permeabilized in a 3% hydrogen peroxide solution (Sinopharm, Shanghai, China) for 15 min after antigen repair. Subsequently, the sections were blocked with 1% bovine serum albumin (BSA) (Sangon Biotechnology, Shanghai, China) for 15 min, and incubated with primary antibodies against HDAC6 (1:100, ABclonal Biotechnology, Wuhan, China) and NLRP3 (1:100, Affinity Biosciences, Changzhou, China) overnight at 4°. On the next day, the sections were incubated with the secondary antibody (1:500, ThermoFisher Scientific, Pittsburgh, PA, USA) at 37° in a humid chamber for 1 h. Afterward, images were observed and captured under a microscope (BX-53, Olympus).

Zhou T., Jia D., Han J., Xu C., You X, & Ge X. (2023). HDAC6 inhibition alleviates acute pulmonary embolism: a possible future therapeutic option. Folia histochemica et cytobiologica, 61(1).

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