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Anti-Crotonyl-Histone H3 (Lys14) Mouse mAb
Catalog Number: PTM-537
$ 435

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications: WB ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat, Monkey

Synonyms: H3K14cr

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 H3K14cr
UniProt ID

P68431

Immunogen
MW (kDa)
Specificity
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat, Monkey
ICC/IF 1:50 Human
ChIP 6 μg/5x106 cells Human
IP 1:25 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 H3 at Lys14 may play a vital role in the epigenetic modulation, including chromatin remodeling and DNA transcriptional regulation.

Cellular location

Nucleus

Images
ELISA

Crotonyl-histone H3 (Lys14) mouse mAb (PTM-537) specificity was determined by
peptide ELISA.
The graph depicts the binding of the antibody to pre-coated crotonylhistone H3 (Lys14) peptide in the presence of increasing concentrations of various
competitor peptides. As shown, only the crotonyl-histone H3 (Lys14) peptide
competed away binding of the antibody.

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: MCF-7, BRL, N2a, COS-7, rH3
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted band size: 17 kDa
Observed band size: 17 kDa

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-537

ChIP

Cell type: MCF7
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/G MagBeads
Description: Chromatin immunoprecipitations were performed with 6 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, RPL30, LDHA, FOXO3a-promoter and RAB20. The data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.

IP

IP of HeLa+Crotonic acid (100ng/ml, 18hr) cells extracts
IP Ab incubation condition: PTM-537, 4°C overnight, 1:25 dilution
WB Primary Ab incubation condition: PTM535, room temperature 2h, 1:2000 dilution
Secondary Ab: Anti-Mouse IgG for IP (HRP Conjugate)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Lane 1: 5% Input
Lane 2: IP with PTM-537 (1:25)
Observed band size: 17 kDa
Exposure time: 60 s

Research Use

For research use only, not for use in diagnostic procedures.

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