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Recombinant Rabbit Monoclonal
Anti-Crotonyl-Histone H3 (Lys4) Rabbit mAb
Catalog Number: PTM-547RM
$ 420

Clone Number: PAPTM-O0301

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K4cr

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 H3K4cr
UniProt ID

P68431

Immunogen Crotonylated human histone H3 (Lys4) peptide
MW (kDa) 15
Specificity Anti-Crotonyl-Histone H3 (Lys4) Rabbit mAb detects histone H3 when it is crotonylated at Lys4
Product Usage Information
Applications Dilution Recommended Species
WB 1:5000 - 1:10000 Human, Rat
IHC-P 1:500 - 1:1000 Human, mouse
IP 1:1000 Human
ChIP 6 μg per 5x106 cells Human
Properties
Purity Protein A 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. Crotonylation of lysine is a newly identified reversible modification controlling chromosome structure and gene transcription. The reversible lysine crotonylation has been well demonstrated in eukaryotic histones from worm to human. The unique structure and genomic localization of histone lysine crotonylation suggest that it is mechanistically and functionally different from histone lysine acetylation. Specifically, in both human somatic and mouse male germ cell genomes, histone crotonylation marks either active promoters or potential enhancers. Crotonylation of histone H3 at Lys4 may play a vital role in the epigenetic modulation, including chromatin remodeling and DNA transcriptional regulation

Cellular location

cellular Localization

Images
Dot Blot

Blocking buffer: 5% NFDM/TBST
Primary ab dilution: 1:50,000
Primary ab incubation condition: 2 hours at room temperature
Secondary ab: Goat Anti-rabbit IgG H&L (HRP)
Peptide amount: 1 ng, 4 ng, 16 ng, 64ng
Exposure time: 30 s
The list of peptides is included in the table below:
WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:10000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: (-): HeLa, (+): HeLa + Crotonic acid (10 mM, 15h)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:10000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: (-): HeLa, (+): HeLa + Sodium Butyrate (5mM, 24hr)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:10000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: (-): C6, (+): C6+ Crotonic acid (10 mM, 15h)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa
IHC-P

Tissue: Human 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 condition: 1 hour at room temperature
Secondary ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use)
Counter stain: Hematoxylin (blue)
Comment: Color brown is the positive signal for PTM-547RM

Tissue: Mouse cerebrum
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 condition: 1 hour at room temperature
Secondary ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use)
Counter stain: Hematoxylin (blue)
Comment: Color brown is the positive signal for PTM-547RM
IP

IP of HeLa+Crotonic acid (10mM 15h) cells extracts:
IP Ab incubation condition: PTM-547RM, 1:100 dilution, 4°C overnight
WB primary Ab incubation condition: PTM-547RM, room temperature 2h , 1:1000 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
1: 10%% Input
2: IP with Rabbit monoclonal IgG Isotype
3: IP with PTM-547RM
Observed MW: 15 kDa
Exposure time: 180 s
CHIP

Cell type: HeLa+Crotonic acid (10mM 15h)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Comment: The ChIP was 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, PRL30, LDHA-promoter, LDHA, FOXO3a promoter, FOXO3a-Downstream, RAB20 and TUBBP10. 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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