Fourth-instar nymphs were treated with dsGfp, dsNlHdac1 or dsNlHdac4, and raised to adults. For immunoblot analysis of H3 and H4 acetylation, ovaries were dissected from females (n = 50) at 0–6 h after eclosion, and total histones were extracted using the EpiQuik total histone extraction kit (Epigentek). For immunoblot analysis of vitellogenin, the fat body was dissected from females (n = 50) at 3 days after eclosion. Equal amounts of protein were loaded for each lane on SDS-PAGE gel. Western blotting was performed with antibodies of acetyl-histone H3 antibody sampler kit (Cell Signaling Technology), histone H3 acetylation antibody panel pack II (Epigentek), acetylhistone H4 antibody sampler kit (Cell Signaling Technology) and anti-vitellogenin mAb. Immuno-reactivity was imaged with the Molecular Imager ChemiDoc XRS+ system (Bio-Rad).
Acetyl histone h3 antibody sampler kit
Manufactured by Cell Signaling Technology
Sourced in United States
The Acetyl-histone H3 antibody sampler kit contains a set of highly specific antibodies that detect various acetylation sites on histone H3. This kit provides a convenient way to analyze the acetylation status of histone H3 in cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Epiquik total histone extraction kit, by Epigentek (1 mentions)
Molecular imager chemidoc xrs system, by Bio-Rad (1 mentions)
Histone h4, by Abcam (1 mentions)
Hrp linked anti rabbit and anti mouse igg, by Cell Signaling Technology (1 mentions)
Anti glial fibrillary acidic protein antibody, by Cell Signaling Technology (1 mentions)
Anti pcna pc10 antibody, by Santa Cruz Biotechnology (1 mentions)
Anti lamin b1 antibody, by Abcam (1 mentions)
Anti flag antibody, by Merck Group (1 mentions)
Anti phospho histone h2a x ser139, by Merck Group (1 mentions)
Anti histone h3 antibody, by Merck Group (1 mentions)
Anti v5 antibody, by Merck Group (1 mentions)
Anti phospho histone h3 ser 10 antibody, by Cell Signaling Technology (1 mentions)
Anti beta tubulin antibody, by Abcam (1 mentions)
4 protocols using acetyl histone h3 antibody sampler kit
1
Histone Acetylation and Vitellogenin Analysis
2
Quantification of Histone Acetylation
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Proteins were extracted from one million cells after 2.5 h of treatment with DMSO or HDACi using RIPA buffer supplemented with 1 M NaF, 10 mM phenylmethylsulfonyl fluoride, and a protease inhibitor cocktail. For detection of specific acetylation of histone H3 and H4, the following antibodies were used: Acetyl-Histone H3 Antibody Sampler kit #9927 (Cell Signaling), anti-H3K23 (D6Y7M, Cell signaling), -H4K5 (EP1000Y, Abcam), -H4K8 (EP1002Y, Abcam), -H4K12 (Abcam), -H4K16 (EPR1004, Abcam), and -Histone H4 (Abcam).
3
Phosphorylated Tau and Histone Acetylation Antibodies
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Monoclonal antibody against phosphorylated tau PHF1 (S396/404), 12E8(S262/356) and against total tau (K9JA) was a kind gift from Prof. Dr. Eckhard Mandelkow and Prof. Dr. Eva-Maria Mandelkow (DZNE, University of Bonn, Germany). Acetyl-Histone H3 antibody sampler kit comprising acetylation of K9, K14, K18, K27, and K56, anti-H4K5ac, and anti-H2BK12ac antibody were from Cell Signaling Technology Danvers, MA, USA (9927, 8647 and 5410). Anti-HDAC1, -HDAC2, -HDAC3, and -HDAC6 antibodies were from Cell Signaling Technology (antibody Sampler Kit #9928). Anti-SGPL1 antibody was from abcam (Cambridge, UK; ab56183) and Anti-glial fibrillary acidic protein (GFAP) antibody from Cell Signaling Technology (12389).Secondary antibodies were HRP linked anti-rabbit and anti-mouse IgG (Cell Signaling Technology, 7074 and 7076). 5,5′-Dimethyl-BAPTA-AM was from Sigma- Aldrich, Munich, Germany (16609).
4
Multiplex Antibody Detection Methodology
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The following antibodies were used: an anti-FLAG antibody (Sigma-Aldrich, F3165, 1:1000); anti-PCNA (PC10) antibody (Santa Cruz Biotechnology, sc-56, 1:2000); Acetyl-Histone H3 Antibody Sampler Kit (Cell Signaling Technology, #9927, 1:2000); Acetyl-Histone H4 Antibody Set (Merck-Millipore, #17-211, 1:2000); anti-histone H3 antibody (Merck-Millipore, #07-690, 1:5000); anti-BRD2 antibody (Bethyl, A302-583A, 1:1000); anti-BRD3 antibody (Bethyl, A302-368A, 1:1000); anti-BRD4 antibody (Bethyl, A301-985A, 1:1000); Pan-BRD4 antibody (Sigma-Aldrich, AV39076, 1:1000); anti-V5 antibody (Sigma-Aldrich, V8137, 1;2000); anti-UAF1 antibody (Santa Cruz Biotechnology, sc-514473, 1:500); anti-S tag antibody (Sigma-Aldrich, SAB2702227, 1:5000); anti-phospho-histone H3 (Ser10) antibody (Cell Signaling Technology, #9706, 1:1000); anti-beta-tubulin antibody (Abcam, ab15568, 1:2000); anti-lamin B1 antibody (Abcam, ab16048, 1:1000); anti-phospho-histone H2A.X (Ser139) (Merck-Millipore, #05-636, 1:2000) . The anti-phospho-S653 ATAD5 antibody (1:1000) and anti-human ATAD5 antibody (1:1000) were raised in rabbits.
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