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Recombinant Rabbit Monoclonal
Anti-Crotonyl-Histone H2B (Lys15/16/20) Rabbit mAb
Catalog Number: PTM-546RM
$ 435

Clone Number: 25-H1L3

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB ChIP

Reactivity: Human

Synonyms: H2BK15/K16/K20cr

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 H2BK15/K16/K20cr
UniProt ID

P62807

Immunogen Crotonylated human histone H2B (Lys15/16/20) peptide
MW (kDa) 14
Specificity Anti-Crotonyl-Histone H2B (Lys15/16/20) Rabbit mAb detects histone H2B when it is crotonylated at Lys15/16/20.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 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. This antibody can simultaneously identify histone H3 at Lys15, 16 and 20.

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: 1-3: 1ng, 4 ng, 16 ng, 64 ng, 4-17: 64 ng
Exposure time: 30 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
Secondary Ab: (-): HeLa, (+): HeLa+ sodium butyrate (30mM, 4hr)
Protein loading quantity: 20 μg
Exposure time: 60 seconds
Predicted band size: 14kDa
Observed band size: 14kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: (-): MCF-7, (+): MCF-7+ sodium butyrate (50mM, 24hr) + trichostatin A (500ng/ml, 4 hr)
Protein loading quantity: 20 μg
Exposure time: 60 seconds
Predicted band size: 14kDa
Observed band size: 14kDa
ChIP

Cell type: MCF-7+Sodium butyrate(50 mM, 24 hr)+trichostatin A (500 ng/ml, 4 hr)
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×1006 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 region, RPL30, LDHA CDS region, 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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