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Anti-Histone H3 (Trimethyl Lys4, Serotonyl Gln5) Rabbit pAb
Catalog Number: PTM-1420
$ 390

Clone Number: /

Host: Rabbit Clonality: Polyclonal

Applications： WB ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K4me3/Q5ser

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 H3K4me3/Q5ser
UniProt ID

P68431

Immunogen Trimethylated/serotonylated human histone H3 (Lys4/Gln5) peptide
MW (kDa) 15
Specificity Anti-Trimethyl/Serotonyl-Histone H3 (Lys4/Gln5) Rabbit pAb detects histone H3 only when it is Trimethylated at Lys4and serotonylated at Gln5.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
ChIP 6 μg/5x106 cells Human
Properties
Purity Protein A 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) are key mechanisms of epigenetics that modulate chromatin structures, termed as “histone code”. Serotonin, also known as 5-hydroxytryptamine (5-HT), is an excitatory neurotransmitter that regulates brain cell activity by activating specific membrane receptors in the brain. The amino group of serotonins can covalently bind to the glutamine residue at the fifth position of histone H3 by TGM2 to form histone 5-hydroxytryptamine (H3Q5ser). H3Q5ser modifies sites adjacent to H3K4me3 form PTM crosstalk. The presence of H3K4me3Q5ser enhanced the binding of transcription factors, such as TFIID to chromatin, thereby activating the expression of downstream genes in developing rodent brains and human neurons.

Cellular location

Nucleus

Images
Dot Blot

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Peptide amount: 1 ng, 4 ng, 16 ng
Exposure time: 60 s
The list of peptides is included in the table below.
WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Lysate: HeLa, N2a, BRL, Mouse kidney
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted band size: 15 kDa
Observed band size: 15 kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: lane 1: PTM-1420 + H3K4me3/Q5ser peptides, lane 2: PTM-1420 + unmodified peptides, lane 3: PTM-1420, 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Lysate: N2a
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted band size: 15 kDa
Observed band size: 15 kDa
ChIP

Cell type: HeLa + sodium lactate (100 mM, 24 hours)
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5x106 cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Description: Chromatin immunoprecipitations were performed with 6 μg of normal rabbit IgG as a negative control. Real time
quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human
GAPDH CDS region, RPL30 Exon 3, LDHA, FOXO3a-promoter, FOXO3a-downstream, RAB20 and
LDHA-promoter. The data are presented as enrichment of each sample relative to total amount of input
chromatin at each amplicon.

Research Use

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

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