Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody [PSH25-67]
Catalog# HA724554
Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody [PSH25-67]
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WB
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IF-Cell
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IHC-P
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IP
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ChIP
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Human
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Mouse
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Rat
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HA751986
不含抗保成分
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unconjugated
Overview
Product Name
Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody [PSH25-67]
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
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:2,000
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IHC-P
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1:500-1:1,000
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IP
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1-2μg/sample
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ChIP
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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.
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
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Histone H4 (acetyl K12) on different lysates with Rabbit anti-Histone H4 (acetyl K12) antibody (HA724554) 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: HA724554, 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: HA724554, 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: HA724554, 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: HA724554, 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: HA724554, 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: HA724554, 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: HA724554, 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 HA724554 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724554 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724554 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724554 in HeLa cell lysate
Exposure time: 2 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724554, 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) (HA724554) / 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"