WB
Cell treatment (CT) Western blot analysis of Histone H4 (acetyl K12) on different lysates with Rabbit anti-Histone H4 (acetyl K12) antibody at 1/50,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 2 seconds; Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary antibody: Rabbit anti-Histone H4 (acetyl K12) antibody, 1/50,000 in primary antibody dilution buffer, overnight at 4 °C. Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.ICC/IF
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: 1/2,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).ICC/IF
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: 1/2,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).ICC/IF
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: 1/2,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).IHC
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: 1/500, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.IHC
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: 1/1,000, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.IHC
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: 1/1,000, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.IP
Immunoprecipitation (IP) Histone H4 (acetyl K12) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody 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. Exposure time: 2 seconds. Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary dilution: 1/5,000 in primary antibody dilution buffer, 2 hours at room temperature.CHIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H4 (acetyl K12) / 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.| Product Name | Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human Histone H4 aa 1-50 (acetyl K12). |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ChIPICC/IFIHCIPWB |
| WB:1:5000 ICC/IF:1:2000 IHC:1:500-1:1000 IP:1-2μg/sample ChIP: Use 0.5~2 μg for 25 μg of chromatin. |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | H4C1 |
|---|---|
| Gene Synonyms | H4FA HIST1H4A |
| Gene Full Name | H4 clustered histone 1 |
| Gene Summary | Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H4 family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6. [provided by RefSeq, Aug 2015] |
| Molecular Weight(MW) | 11 kDa(Observed band size: 15 kDa) |
| Cellular Localization | Nucleus. Chromosome. |
| SwissProt ID | P62805 |
|---|
WB
Cell treatment (CT) Western blot analysis of Histone H4 (acetyl K12) on different lysates with Rabbit anti-Histone H4 (acetyl K12) antibody at 1/50,000 dilution. Lysates/proteins at 10 µg/Lane. Exposure time: 2 seconds; Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary antibody: Rabbit anti-Histone H4 (acetyl K12) antibody, 1/50,000 in primary antibody dilution buffer, overnight at 4 °C. Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.
ICC/IF
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: 1/2,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
ICC/IF
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: 1/2,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
ICC/IF
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: 1/2,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
IHC
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: 1/500, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.
IHC
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: 1/1,000, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.
IHC
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: 1/1,000, 1 hour at room temperature. Secondary antibody: 20 minutes at room temperature.
IP
Immunoprecipitation (IP) Histone H4 (acetyl K12) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Histone H4 (acetyl K12) Recombinant Rabbit Monoclonal Antibody 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. Exposure time: 2 seconds. Blocking: 5% NFDM/TBST, 1 hour at room temperature. Primary dilution: 1/5,000 in primary antibody dilution buffer, 2 hours at room temperature.
CHIP
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H4 (acetyl K12) / 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.| Application Notes | WB:1:5000 ICC/IF:1:2000 IHC:1:500-1:1000 IP:1-2μg/sample ChIP: Use 0.5~2 μg for 25 μg of chromatin. |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
