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Recombinant Rabbit Monoclonal
Anti-L-Lactyl-Histone H3 (Lys27) Rabbit mAb
Catalog Number: PTM-1428RM
$ 420

Clone Number: JMMR-P2629

Host: Recombinant Monoclonal Clonality: Recombinant Monoclonal

Applications： WB IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H3K27la

Product Size
100μl
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 H3K27la
UniProt ID

P68431

Immunogen L-lactylated human histone H3 (Lys27) peptide
MW (kDa) 15
Specificity Anti-L-Lactyl-Histone H3 (Lys27) Rabbit mAb detects histone H3 only when it is L-lactylated at Lys27.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000-1:5000 All
IP 1:100-1:500 Human
ChIP 6 μg per 5×106 cells Human
Properties
Purity Protein A 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, 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

Blocking buffer: 5% NFDM/TBST
Primary ab dilution: 1:50000
Primary ab incubation condition: 2 hours at room temperature
Secondary ab: Goat Anti-rabbit IgG H&L (HRP)
Peptide amount: Lane1, Land2 and Lane3: 1 ng, 4 ng, 16 ng, 64ng; other spots: 64ng
Exposure time: 60 s
Note: Lane2 and Lane3 is two different peptides to H3K27hib.
WB

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:20000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: (-): HeLa, (+): HeLa+ Sodium Lactate (25mM, 24h)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:20000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: (-): Neuro-2a, (+): Neuro-2a+ Sodium Lactate (25mM, 24h)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa

Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:20000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: (-): PC-12, (+): PC-12+ Sodium Lactate (25mM, 24h)
Protein loading amount: 20 μg
Exposure time: 60 s
Predicted MW: 15 kDa
Observed MW: 15 kDa
IP

IP of HeLa+Lac Na(25mM, 24h) cells extracts:
IP Ab incubation condition: PTM-1428RM, 4°C overnight, 1:50 dilution
WB primary Ab incubation condition: PTM-1428RM, room temperature 2h , 1:500 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
1: 2.5% Input
2: IP with PTM-1428RM
3: IP with Rabbit monoclonal IgG Isotype
Observed MW: 15 kDa
Exposure time: 30 s

IP of HeLa cells extracts:
IP Ab incubation condition: PTM-1428RM, 4°C overnight, 1:50 dilution
WB primary Ab incubation condition: PTM-1428RM, room temperature 2h , 1:500 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
1: 2.5% Input
2: IP with PTM-1428RM
3: IP with Rabbit monoclonal IgG Isotype
Observed MW: 15 kDa
Exposure time: 30 s
ChIP

Cell type: HeLa+ sodium lactate (25 mM, 24 hr)
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
Comment: The ChIP was performed with 6 μg of normal Rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human GAPDH-CDS, RPL30, TuBBP10 and Foxo3a-D. Data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon.

Research Use

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

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