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Recombinant Rabbit Monoclonal
Anti-L-Lactyl-Histone H3 (Lys14) Rabbit mAb
Catalog Number: PTM-1414RM
$ 420

Clone Number: PA-111-9

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB ICC/IF ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K14la

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 H3K14la
UniProt ID

P68431

Immunogen L-lactylated human histone H3 (Lys14) peptide
MW (kDa) 15
Specificity Anti-L-Lactyl-Histone H3 (Lys14) Rabbit mAb detects endogenous levels of histone H3 when it is lactylated at Lys14.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 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

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: 1 ng, 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: H3K14 L-lactyl. Lane 2: H3K23 L-lactyl.
Lane 3: H3K14 crotonyl. Lane 4: H3K14 acetyl.
Lane 5: H3K14 propionyl. Dot 6: H3K23 succinyl.
Dot 7: H3K14 2-hydroxyisobutyryl. Dot 8: H3K14 β-hydroxybutyryl.
Dot 9: H3K14 monomethyl. Dot 10: H3K14 Dimethyl.
Dot 11: H3K27 L-lactyl. Dot 12: H3K9 L-lactyl.
Dot 13: H4K8 acetyl. Dot 14: H4K8 2-hydroxyisobutyryl.
Dot 15: H4K8 benzoyl. Dot 16: H4K8 L-lactyl.
Dot 17: H4K8 crotonyl. Dot 18: H3K14 unmodified.
WB

Lysates: (-): HeLa cells; (+): HeLa cells treated with 100 mM sodium lactate for 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: 15 kDa
Observed band size: 15 kDa

Samples: 1. Neuro-2a; 2. BRL; 3. Rat kidney; 4. Mouse kidney; 5. Recombinant histone H3 (20 ng)
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: 15 kDa
Observed band size: 15 kDa
ICC/IF

Samples: (A) HeLa cells; (B) HeLa cells treated with 100 mM sodium lactate for 24 hours
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:100
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-1414RM.
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 CDDS, 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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