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Anti-Crotonyl-Histone H2B (Lys12) Rabbit pAb
Catalog Number: PTM-509
$ 435

Clone Number: /

Host: Rabbit Clonality: Polyclonal

Applications： WB ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H2BK12cr

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 H2BK12cr
UniProt ID

P62807

Immunogen
MW (kDa) 14
Specificity
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:1000 Human, Mouse, Rat
ChIP 12 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

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 Lys12 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: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Immunogen peptide quantity: 1 ng, 4 ng, 16 ng
Exposure time: 15 s
The list of peptides is included in the table below.
WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Lysate: (-) HeLa, (+) HeLa+Sodium crotonylate (10mM, 4hr)
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 14 kDa
Observed band size: 14 kDa
ChIP

Cell type: HeLa treated with Crotonic acid (10mM 15h)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 12 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Description: Chromatin prepared from 5 x 106 HeLa treated with Crotonic acid (10mM 15h) was subjected to chromatin immunoprecipitation (ChIP) using either serial dilutions of AntiCrotonyl-Histone H2B (Lys12) Rabbit pAb or 1 μg of normal rabbit IgG as a negative control.
The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human LDHA, FOXO3a-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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References (3)
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    Year 2022
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    Authors Xiao-Feng Chen, et al.
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  • Chromodomain Protein CDYL Acts as a Crotonyl-CoA Hydratase to Regulate Histone Crotonylation and Spermatogenesis
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    Authors Shumeng Liu, et al.
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