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Recombinant Rabbit Monoclonal
Anti-Crotonyl-Histone H3 (Lys27) Rabbit mAb
Catalog Number: PTM-545RM
$ 435

Clone Number: PFPTM-546-27

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K27cr

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 H3K27cr
UniProt ID

P68431

Immunogen Crotonylated human histone H3 (Lys27) peptide
MW (kDa) 15
Specificity Anti-Crotonyl-Histone H3 (Lys27) Rabbit mAb detects histone H3 when it is crotonylated at Lys27.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000 - 1:2000 Human, Mouse, Rat
ICC/IF 1:50 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

Lysine crotonylation is a newly-identified histone post-translational modification that has come to light through integrated proteomic approaches and elaborate biochemistry analyses. Lysine crotonylation has been found to be evolutionarily conserved across multiple species, spanning both prokaryotes and eukaryotes, and occurs in a wide range of proteins, incuding histones and non-histone substrates. The unique structure and genomic localization of lysine crotonylation suggest that it is mechanistically and functionally different from lysine acetylation. Given the revealed roles of lysine crotonylation in spermatogenesis, the novel modification may play important roles in multiple cellular pathophysiological processes by exerting its effects on chromatin structure and epigenetic pathways.

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 1ng, 4 ng, 16 ng, 64 ng
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)
Exposure time: 30 seconds
The list of peptides used in the experiment is provided below.
Lane 1: H3K27 crotonyl. Lane 2: H3K27 butyryl. Lane 3: H3K9 crotonyl.
Dot 4: H3K27 acetyl. Dot 5: H3K27 2-hydroxyisobutyryl. Dot 6: H3K9 butyryl.
Dot 7: H3K27 β-hydroxybutyryl. Dot 8: H3K9 acetyl. Dot 9: H3K9 crotonyl.
Dot 10: H3K9 β-hydroxybutyryl. Dot 11: H3K9 2-hydroxyisobutyryl.
Dot 12: H3K23 crotonyl. Dot 13: H3K27 unmodified.
WB

Lysates: (-): HeLa cells; (+): HeLa cells treated with 10 mM crotonic acid for 15 hours
Protein loading amount: 20 μg
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)
Exposure time: 30 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa

Lysates: (-): 293T cells; (+): 293T cells treated with 10 mM crotonic acid for 15 hours
Protein loading amount: 20 μg
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)
Exposure time: 30 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
ICC/IF

Samples: (A) HeLa cells; (B) HeLa cells + crotonic acid (10 mM, 15 hours)
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:50
Primary Ab 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-545RM.
ChIP

Sample: HeLa cells + crotonic acid （10 mM, 15 hours)
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 MagBeads
Description: Chromatin immunoprecipitations were performed with 6 μg of normal rabbit IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH CDS, RPL30, LDHA-promoter, LDHA CDS, FOXO3a-promoter, FOXO3a-downstream, RAB20, and TUBBP10 regions. The data are presented as enrichment of each sample relative to total amount of input at each amplicon.

Research Use

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

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