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Recombinant Rabbit Monoclonal
Anti-L-Lactyl-Histone H3 (Lys18) Rabbit mAb (ChIP Grade)
Catalog Number: PTM-1427RM
$ 435

Clone Number: PAPTM-599-16

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB ChIP CUT&Tag

Reactivity: Human, Mouse

Synonyms: H3K18la

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 H3K18la
UniProt ID

P68431

Immunogen L-lactylated human histone H3 (Lys18) peptide
MW (kDa) 15
Specificity Anti-L-Lactyl-Histone H3 (Lys18) Rabbit mAb (ChIP Grade) detects histone H3 only when it is L-lactylated at Lys18.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:1000 Human, Mouse
ChIP 6 μg per 5×106 cells Human, Mouse
CUT&Tag 1:100 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, a recently discovered post-translational modification induced by lactate has emerged as a significant addition to this repertoire. The extent and dynamics of this modification are highly reliant on lactate levels within the cellular microenvironment and can be modulated through the introduction of extracellular lactate in cultured cells or the stimulation of intracellular glycolysis. 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). Histone lactylation has been implicated in various biological processes, including inflammation, fibrosis, differentiation, and cancer progression.

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 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: 10 seconds
The list of peptides used in the experiment is provided below.
Lane 1: H3K18 L-lactyl. Lane 2: H3K18 acetyl.
Lane 3: H3K18 crotonyl. Lane 4: H3K18 butyryl.
Lane 5: H3K18 propionyl. Lane 6: H3K18 2-hydroxyisobutyryl.
Lane 7: H3K18 β-hydroxybutyryl. Lane 8: H3K18 unmodified
WB

Lysates: (-): HeLa cells; (+): HeLa cells treated with 25 mM sodium lactate for 24 hours
Protein loading amount: 20 μg
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)
Exposure time: 60 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa

Lysates: (-): K562 cells; (+): K562 cells treated with 25 mM sodium lactate for 24 hours
Protein loading amount: 20 μg
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)
Exposure time: 60 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa

Lysates: (-): NIH/3T3 cells; (+): NIH/3T3 cells treated with 25 mM sodium lactate for 24 hours
Protein loading amount: 20 μg
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)
Exposure time: 60 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
ChIP

Sample: Hela cells treated with 100 mM sodium lactate for 24 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, LDHA-promoter, LDHA, FOXO3a-downstream, RAB20, and TUBBP10 regions. The data are presented as enrichment of each sample relative to total amount of input at each amplicon.
CUT&Tag

Sample: HeLa cells treated with 100 mM sodium lactate for 24 hours
Cell quantity: 1×105 cells
Primary Ab dilution: 1:100
Primary Ab incubation condition: 4⁰C overnight
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (PTM-6252)
Secondary Ab dilution: 1:1400
Description: CUT&Tag was performed with Anti-L-Lactyl-Histone H3 (Lys18) Rabbit mAb (PTM-1427RM) alongside normal rabbit IgG as a negative control. The resulting DNA was sequenced on the Illumina NovaSeq platform with paired-end reads of 150 bp and a sequencing depth of 3 million reads. The figure shows significant H3K18la signal enrichments at the promoter regions of CD9, PLEKHG6, LTBR, and GAPDH.

Research Use

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

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