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Anti-Butyryl-Histone H4 (Lys5) Mouse mAb
Catalog Number: PTM-310
$ 420

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications： WB IHC-P ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H4K5bu

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 H4K5bu
UniProt ID

P62805

Immunogen Butyrylated human histone H4 (Lys5) peptide
MW (kDa) 11
Specificity Anti-Butyryl-Histone H4 (Lys5) Mouse mAb detects histone H4 only when it is butyrylated at Lys5. 
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ChIP 6 μg per 5x106 cells 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

Histones are subject to a variety of enzyme catalyzed modifications, including acetylation, methylation, phosphorylation, ubiquitylation, etc. Butyrylation of lysine, structurally similar to lysine acetylation, is a newly identified reversible modification controlling protein activity. The reversible lysine butyrylation has been well demonstrated in both prokaryotes and eukaryotes in wide ranges of proteins including histones and non-histone substrates, such as p53. Similar to acetylation of histione H4 at lysine 12, butyrylation of histone H4 at lysine 5 may play a vital role in the epigenetic modulation, including chromatin remodeling and DNA transcriptional regulation.

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: 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: H4K5 butyryl. Lane 2: H4K5 acetyl.
Lane 3: H4K5 2-hydroxyisobutyryl. Lane 4: H4K5 β-hydroxybutyryl.
Lane 5: H4K5 crotonyl. Lane 6: H4K8 butyryl.
Lane 7: H4K8 2-hydroxyisobutyryl. Lane 8: H4K8 β-hydroxybutyryl.
Lane 9: H4K8 crotonyl. Lane 10: H4K12 propionyl.
Lane 11: H4K12 butyryl. Lane 12: H4K12 succinyl.
Lane 13: H4K12 crotonyl. Lane 14: H4K5 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: Human spleen
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: 1:200
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-310.

Tissue: Mouse testis
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-310.

Tissue: Rat spleen
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-310.
ChIP

Sample: HeLa cells, serum starvation for 12 hours, followed by incubation with 5 mM sodium butyrate for 24 hours.
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 10 μg
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 promoter, GAPDH CDS, RPL30 exon 3, FOXO3a promoter, and FOXO3a downstream regions. The data are presented as enrichment of each sample relative to total amount of input at each amplicon.

Research Use

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

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    Anti-Butyryl-Histone H4 (Lys5) Rabbit pAb
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    Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
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References (2)
  • Butyrylation Meets Adipogenesis-Probed by a p300-Catalyzed Acylation-Specific Small Molecule Inhibitor: Implication in Anti-obesity Therapy
    Year 2022
    Journal JOURNAL OF MEDICINAL CHEMISTRY
    Authors Aditya Bhattacharya, et al.
    Applications Unspecified
    Reactivity
  • Metabolically controlled histone H4K5 acylation/acetylation ratio drives BRD4 genomic distribution
    Year 2021
    Journal Cell Reports
    Authors Mengqing Gao, et al.
    Applications Unspecified
    Reactivity
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