Histone H4 [ac Lys12] Antibody - BSA Free

Novus Biologicals | Catalog # NBP2-59266

Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse

Applications

Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing, Dot Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
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Product Specifications

Immunogen

The exact immunogen is proprietary information.

Modification

ac Lys12

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Histone H4 [ac Lys12] Antibody - BSA Free

Western Blot: Histone H4 [ac Lys12] Antibody [NBP2-59266]

Western Blot: Histone H4 [ac Lys12] Antibody [NBP2-59266]

Western Blot: Histone H4 [ac Lys12] Antibody [NBP2-59266] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H4K12ac. The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right, the marker (in kDa) is shown on the left.
Immunocytochemistry/ Immunofluorescence: Histone H4 [ac Lys12] Antibody [NBP2-59266]

Immunocytochemistry/ Immunofluorescence: Histone H4 [ac Lys12] Antibody [NBP2-59266]

Immunocytochemistry/Immunofluorescence: Histone H4 [ac Lys12] Antibody [NBP2-59266] - HeLa cells were stained with the antibody against H4K12ac and with DAPI. Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Chromatin Immunoprecipitation: Histone H4 [ac Lys12] Antibody [NBP2-59266] - ChIP assays were performed using human HeLa cells, the antibody against H4K12ac and optimized PCR primer sets for qPCR. ChIP was performed with the ChIP-seq kit on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ELISA: Histone H4 [ac Lys12] Antibody [NBP2-59266]

ELISA: Histone H4 [ac Lys12] Antibody [NBP2-59266]

ELISA: Histone H4 [ac Lys12] Antibody [NBP2-59266] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H4K12ac in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:14,300.
Dot Blot: Histone H4 [ac Lys12] Antibody [NBP2-59266] - To test the cross reactivity of the antibody against H4K12ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac.

Applications for Histone H4 [ac Lys12] Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

0.5-1 ug

Dot Blot

1:1000

ELISA

1:1000

Immunocytochemistry/ Immunofluorescence

1:200

Western Blot

1:500

Formulation, Preparation, and Storage

Purification

Peptide affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide and 0.05% ProClin 300

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: Histone H4

Histone proteins H3, H4, H2A, and H2B function as building blocks to package eukaryotic DNA into repeating nucleosome units that are folded in higher order chromatin fibers. The nucleosome is composed of an octamer containing a H3/H4 tetramer and two H2A/H2B dimers, surrounded by approximately 146 base pairs of DNA. A diverse and elaborate array of post-translational modifications including acetylation, phosphorylation, methylation, ubiquitination, and ADP-ribosylation occurs on the N-terminal tail domains of histones. Methylation of position-specific lysine residues in histone N termini is a central modification for regulating epigenetic transitions in chromatin. Each methylatable lysine residue can exist in a mono, di, or tri methylated state. Arginine resdiues can also by mono or di methylated.

Alternate Names

H4, H4/M, H4FM, H4M, HIST1H4I, HIST4H4, Histone Cluster 1 H4, Histone Cluster 1 H4i, Histone Cluster 4 H4, H4K12ac

Gene Symbol

H4C16

Additional Histone H4 Products

Product Documents for Histone H4 [ac Lys12] Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for Histone H4 [ac Lys12] Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

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Protocols

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Associated Pathways

Notch Signaling Pathways Notch Signaling Pathway Thumbnail