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Recombinant Rabbit Monoclonal
Anti-L-Lactyl-Histone H4 (Lys16) Rabbit mAb
Catalog Number: PTM-1417RM
$ 435

Clone Number: 2H2L1

Host: Rabbit Clonality: Recombinant Monoclonal

Applications: WB IHC-P ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H4K16la

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 H4K16la
UniProt ID

P62805

Immunogen L-lactylated human histone H4 (Lys16) peptide
MW (kDa) 11
Specificity Anti-L-Lactyl-Histone H4 (Lys16) Rabbit mAb detects histone H4 only when it is L-lactylated at Lys16.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:100 - 1:500 Human
ICC/IF 1:500 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, fundamental proteins involved in chromatin structure and gene regulation, are subject to a wide array of enzyme-catalyzed modifications, including acetylation, methylation, phosphorylation, ubiquitination, and numerous others. Histone L-lactylation is a recently discovered post-translational modification that is induced by lactate, a metabolite produced during glycolysis. Regulated by glucose metabolic dynamics and lactate levels within the cellular microenvironment, histone lactylation is implicated in various biological processes, including inflammation, fibrosis, differentiation, and cancer progression. The introduction of lysine L-lactylation is mediated by the acetyltransferase p300, while the removal of lactylation marks from histones has been attributed to Class I histone deacetylases (HDAC 1-3). Notably, Sirtuin 3 (SIRT3) has been reported as an eraser of H4K16 lactylation (H4K16la).

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 1 ng, 4 ng, 16 ng
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:10000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 10 seconds
The list of peptides used in the experiment is provided in the table below.

WB

Lysates: (-): HeLa cells; (+): HeLa cells + sodium lactate (100 mM, 24 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: 60 seconds
Predicted band size: 11 kDa
Observed band size: 11 kDa

IHC-P

Tissue: Human colon cancer
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: 1:500
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-1417RM.

ICC/IF

Samples: (A) HeLa cells; (B) HeLa cells + sodium lactate (100 mM, 24 hours)
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:500
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-1417RM.

ChIP

Sample: HeLa cells + sodium lactate (100 mM, 24 hours)
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5x106 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 RPL30 Exon 3 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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References (1)
  • Comprehensive Analysis of Lysine Lactylation in Rice (Oryza sativa) Grains
    Year 2021
    Journal JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
    Authors Xiaoxi Meng, et al.
    Applications Unspecified
    Reactivity