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Histone H4 (acetyl K8) Recombinant Rabbit Monoclonal Antibody [PS01-25]

Usd: 385

Specification

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Catalog# HA721249

Histone H4 (acetyl K8) Recombinant Rabbit Monoclonal Antibody [PS01-25]

Application

  • WB

  • IF-Cell

  • IHC-P

  • ChIP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

Histone H4 (acetyl K8) Recombinant Rabbit Monoclonal Antibody [PS01-25]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Histone H4 aa 1-20 (acetyl K8).

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, ChIP

Target Molecular Weight

Predicted band size: 11 kDa

Positive Control

Human skin tissue, rat skin tissue, human colon carcinoma tissue, PC-12, HeLa (Human cervical adenocarcinoma cell) cell lysate, NIH/3T3 (Human embryonic kidney cell) cell lysate.

Conjugation

unconjugated

Clone Number

PS01-25

RRID

AB_3072368

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5000

  • IF-Cell

  • 1:50

  • IHC-P 1

  • 1,000-1:5,000

  • ChIP

  • Use 0.5~2 μg for 25 μg of chromatin.

Target

Function

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. H4K8ac is part of 17 modifications of a group of active promoters. H4K8ac is found more often in active promoters and transcribed regions than other marks. H4K8ac is modified by a different group of enzymes than other H4 lysines.

Background References

1. Huang, Suming; Litt, Michael D.; Ann Blakey, C. (2015-11-30). Epigenetic Gene Expression and Regulation. pp. 21–38.

2. Sadoul K, Boyault C, Pabion M, Khochbin S (2008). "Regulation of protein turnover by acetyltransferases and deacetylases". Biochimie. 90 (2): 306–12.

Subcellular Location

Nucleus. Chromosome.

UNIPROT

SwissProt: P62805 Human

SwissProt: P62806 Mouse

SwissProt: P62804 Rat

Synonyms

dJ160A22.1 antibody

dJ160A22.2 antibody

dJ221C16.1 antibody

dJ221C16.9 antibody

FO108 antibody

H4 antibody

H4.k antibody

H4/a antibody

H4/b antibody

H4/c antibody

Expand

Images

  • Western blot analysis of Histone H4 (acetyl K8) on different lysates with Rabbit anti-Histone H4 (acetyl K8) antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate<br />Lane 2: NIH/3T3 (Human embryonic kidney cell) cell lysate<br /><br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 46 seconds; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 11 kDa<br />Observed band size: 11 kDa

    Western blot analysis of Histone H4 (acetyl K8) on different lysates with Rabbit anti-Histone H4 (acetyl K8) antibody (HA721249) at 1/5,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
    Lane 2: NIH/3T3 (Human embryonic kidney cell) cell lysate


    Lysates/proteins at 15 µg/Lane.
    Exposure time: 46 seconds; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA721249, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 11 kDa
    Observed band size: 11 kDa

  • Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H4 (acetyl K8) antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Histone H4 (acetyl K8) antibody (HA721249) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721249) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-Histone H4 (acetyl K8) antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-Histone H4 (acetyl K8) antibody (HA721249) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721249) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Histone H4 (acetyl K8) antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Histone H4 (acetyl K8) antibody (HA721249) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721249) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of PC-12 cells labeling Histone H4 (acetyl K8) with Rabbit anti-Histone H4 (acetyl K8) antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H4 (acetyl K8) antibody (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of PC-12 cells labeling Histone H4 (acetyl K8) with Rabbit anti-Histone H4 (acetyl K8) antibody (HA721249) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Histone H4 (acetyl K8) antibody (HA721249) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

  • Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H4 (acetyl K8) (<a href="/products/HA721249" style="font-weight: bold;text-decoration: underline;">HA721249</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H4 (acetyl K8) (HA721249) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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