Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Biochemical basis of the HDAC activity assay in brain and other tissues. Figure created using BioRender.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: HDAC Activity Assay

Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Illustration of critical steps involved in measuring HDAC activity in the mice tissues. Figure created using BioRender.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: Activity Assay

Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Assay layouts for protein estimation and HDAC activity. ( A ) Representative sample layout of the 96‐well microplate for the BCA assay. ( B ) Representative sample layout of the 96‐well microplate for the HDAC activity assay. ( C ) A typical standard curve generated using the Spectra MAX Gemini XS Plate Reader is shown with standard concentrations of 50, 250, 500, 1000, 1500, and 2000 µg/ml at an excitation wavelength of 360 nm and emission wavelength of 460 nm. Panels A and B were created using BioRender.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: Activity Assay, BIA-KA, HDAC Activity Assay, Generated

Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Quantification of HDAC activity in brain and other tissues. ( A ) Basal HDAC activity in sham brain tissues 7 days after TBI. The spleen is shown to have the highest normalized HDAC activity. ( B ) Percentage HDAC activity normalized to hippocampus levels. ( C and D ) Inhibition of HDAC activity by HDAC inhibitors 24 hr after TBI in the hippocampus of male mice is shown as ( C ) normalized HDAC activity and ( D ) percentage change in HDAC activity. ( E ) Normalized HDAC activity in the cortex and hippocampus of male and female mice 24 hr after TBI. Data were normalized to samples with trichostatin A (TSA) and are presented as normalized HDAC activity. TBI, traumatic brain injury; ROMI, romidepsin; SAHA, vorinostat; SB, sodium butyrate; FEU, fluorescence emission unit. Data represent mean ± S.E.M. ( n = 6 to 8 for each group). In subpanels C and D, * p < .05 versus sham; # p < .05 versus TBI. In subpanel E, * p < .05 versus sham (same sex); # p < .05 versus TBI (across sex); & p < .05 versus sham (across sex). Data were analyzed using one‐way ANOVA followed by Tukey's post hoc test.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: Activity Assay, Inhibition, Fluorescence