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Anti-Acetyl-Histone H4 (Lys8) Mouse mAb
Catalog Number: PTM-164
$ 340

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications： WB IHC-P ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H4K8ac

Product Size
100 μl
Quantity

Shipping: Ambient temperature

Order online or send purchase order to info@ptmbio.com

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms H4K8ac
UniProt ID

P62805

Immunogen Acetylated human histone H4 (Lys8) peptide
MW (kDa) 11
Specificity Anti-Acetyl-Histone H4 (Lys8) Mouse mAb detects histone H4 only when it is acetylated at Lys8.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Mouse
ICC/IF 1:50 - 1:200 Human
ChIP 1:25 Human
Properties
Purity Protein G and immunogen affinity purified
Constituents PBS, Glycerol, BSA
Storage Store at -20°C. Avoid freeze/thaw cycles.
Stability Stable for 12 months from date of receipt/reconstitution.
Target Information

Background

Histone post-translational modifications (PTMs), known as the “histone code”, are key mechanisms of epigenetics that modulate chromatin structures. The PTMs on histone including acetylation, methylation, phosphorylation, and novel acylations directly affect the accessibility of chromatin to transcription factors and other epigenetic regulators, altering genome stability and gene transcription. Histone acetylation, tightly controlled by the opposing action of histone acetyltransferases (HATs) and histone deacetylases (HDACs), occurs primarily at lysine residues on the N-terminal tails of histones H2A (Lys5, 9, and 15), H2B (Lys5, 12, 15, 16, and 20), H3 (Lys4, 9, 14, 18, 23, 27, and 36), and H4 (Lys5, 8, 12, 16, and 20), and plays vital roles in the regulation of gene expression, DNA damage repair, chromatin dynamics, etc.

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 1 ng, 4 ng, 16 ng
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
The list of peptides used in the experiment is provided below.
Lane 1: H4K8 acetyl. Lane 2: H4K5 acetyl.
Lane 3: H4K5 butyryl. Lane 4: H4K5 crotonyl.
Lane 5: H4K8 butyryl. Lane 6: H4K8 crotonyl.
Lane 7: H4K12 acetyl. Lane 8: H4K12 propionyl.
Lane 9: H4K12 butyryl. Lane 10: H4K12 crotonyl.
Lane 11: H4K8 unmodified.
WB

Lysates: (-) HeLa cells; (+) HeLa cells treated with 30 mM sodium butyrate for 4 hours
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 11 kDa
Observed band size: 11 kDa
IHC-P

Tissue: Mouse colon
Section type: Formalin-fixed & paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:1000
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-164.
ICC/IF

Samples: HeLa cells
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:200
Primary Ab incubation: 4°C overnight
Secondary Ab: Goat Anti-Mouse IgG
Nuclear counter stain: DAPI (blue)
Counter stain: Tubulin (red)
Description: The green color represents the positive signal observed with PTM-164.
ChIP

Sample: HeLa cells
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 1:25 (2 μL)
Beads type and amount per IP: 50 μL of Protein A/G MagBeads
Description: Chromatin immunoprecipitations were performed with 1 μg of normal mouse IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH CDS, GAPDH promoter, RPL30 exon 3, and MyoD1 exon 1 regions. The data are presented as enrichment of each sample relative to the total amount of input chromatin at each amplicon.

Research Use

For research use only, not for use in diagnostic procedures.

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References (8)
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