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Anti-Crotonyl-Histone H2B (Lys16) Mouse mAb
Catalog Number: PTM-533
$ 420

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications： WB IHC-P ICC/IF ChIP

Reactivity: Human, Mouse, Rat, Monkey

Synonyms: H2BK16cr

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 H2BK16cr
UniProt ID

P62807

Immunogen
MW (kDa) 14
Specificity
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat, Monkey
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ICC/IF 1:50 Human
ChIP 12 μg/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. 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 H2B at Lys16 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: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
Immunogen peptide quantity: 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-Mouse IgG H&L pAb (HRP Conjugate)
Lysate: HeLa, C2C12, COS-7, rH2B
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 14 kDa
Observed band size: 14 kDa
IHC-P

Tissue: Human placenta
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)
Description: The brown color represents the positive signal observed with PTM-533

Tissue: Mouse brain
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)
Description: The brown color represents the positive signal observed with PTM-533

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 condition: 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-533
ICC/IF

Cell line: (A) HeLa, (B) HeLa+Crotonic acid (100ng/ml, 18hr)
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% TritonX-100
Primary Ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Counter stain: Tubulin (Red)
Description: The green color represents the positive signal observed with PTM-533
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: 12 ug
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 region, RAB20, RPL30 Exon 3, LDHA, FOXO3a-promoter and FOXO3adownstream. 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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