Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody [PSH25-67]

Overview

Product Name

Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody [PSH25-67] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Histone H4 aa 1-50 (acetyl K12).

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IP, ChIP

Target Molecular Weight

Predicted band size: 11 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cell) cell lysate, HeLa treated with 1μM TSA for 24 hours cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, NIH/3T3 treated with 400nM TSA for 18 hours cell lysate, C6 (Rat glioma cell) cell lysate, C6 treated with 1μM TSA for 18 hours cell lysate.

Conjugation

unconjugated

Clone Number

PSH25-67

Product Features

Form

Liquid

Concentration

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

Lot Concentration Lookup

Storage Instructions

Store at 2-8℃. Avoid freeze.

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:5,000

IF-Cell
1:2,000

IHC-P
1:500-1:1,000

IP
1-2μg/sample

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

☑ Cell treatment (CT)

Western blot analysis of Histone H4 (acetyl K12) on different lysates with Rabbit anti-Histone H4 (acetyl K12) antibody (HA751986) at 1/50,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HeLa treated with 1μM TSA for 24 hours cell lysate
Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate
Lane 5: C6 (Rat glioma cell) cell lysate
Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 2 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751986, 1/50,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
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: 15 kDa
Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: HeLa (Human cervical adenocarcinoma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751986, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblast)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751986, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: C6 (Rat glioma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751986, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Brain
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751986, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751986, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA751986, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Immunoprecipitation (IP)

Histone H4 (acetyl K12) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA751986 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751986 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA751986 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA751986 in HeLa cell lysate

Exposure time: 2 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA751986, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 11 kDa
Observed band size: 15 kDa
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H4 (acetyl K12) (HA751986) / Competitor's antibody / 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.