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Recombinant Rabbit Monoclonal
Anti-Crotonyl-histone H3 (Lys9) Rabbit mAb
Catalog Number: PTM-516RM
$ 420

Clone Number: 2H2L1

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K9cr

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 H3K9cr
UniProt ID

P68431

Immunogen Crotonylated human histone H3 (Lys9) peptide
MW (kDa) 15
Specificity Anti-Crotonyl-histone H3 (Lys9) Rabbit mAb detects histone H3 only when it is Crotonylated at Lys9.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ICC/IF 1:50 Human
IP 1:50 - 1:100 Human
ChIP 12 μg/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 Lys9 may play a vital role in the epigenetic modulation, including chromatin remodeling and DNA transcriptional regulation.

Cellular location

Nucleus

Images
Dot Blot

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1: 10000
Primary Ab incubation condition: 2 hours at room temperature
Immunogen peptide quantity: 4 ng, 16 ng, 64 ng
Exposure time: 60 seconds
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
Lysate: (-) HeLa, (+)HeLa+crotonic acid (100 ng/ml, 18hr)
Protein loading quantity: 20 μg
Exposure time: 60 seconds
Predicted band size: 17 kDa
Observed band size: 17 kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1: 2000
Primary Ab incubation condition: 2 hours at room temperature
Lysate: N2a
Protein loading quantity: 20 μg
Exposure time: 60 seconds
Predicted band size: 17 kDa
Observed band size: 17 kDa
IHC-P

Tissue: Human liver cancer
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
Counter stain: Hematoxylin
Description: The brown color represents the positive signal observed with PTM-516RM

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 condition: 1 hour at room temperature
Counter stain: Hematoxylin
Description: The brown color represents the positive signal observed with PTM-516RM

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 condition: 1 hour at room temperature
Counter stain: Hematoxylin
Description: The brown color represents the positive signal observed with PTM-516RM

Tissue: Rat 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 condition: 1 hour at room temperature
Counter stain: Hematoxylin
Description: The brown color represents the positive signal observed with PTM-516RM
ICC/IF

Cell line: (A) HeLa, (B) HeLa + sodium butyrate (30mM, 4hr)
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:50
Primary incubation condition: 4°C overnight
Nuclear counter stain: DAPI (Blue)
Description: The green color represents the positive signal observed with PTM-516RM
IP

IP of HeLa+ sodium butyrate (30mM, 4hr) cells extracts
IP Ab incubation condition: PTM-113RM, 4°C overnight, 1:100
WB Primary Ab incubation condition: PTM-113RM, room temperature 2h, 1:2000 Secondary Ab: Anti-Rabbit IgG for IP (HRP Conjugate)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
1: 5% Input
2: IP with PTM-113RM
3: IP with Rabbit monoclonal IgG Isotype
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 60 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: 12ug
Beads type and amount per IP: 50 μl of Protein A/G MagBeads
Comment: The ChIP was performed with 1 μg of normal rabbit IgG as a negative control.
Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human RAB20, RPL30, LDHA, FOXO3a-promoter and FOXO3a-downstream. 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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