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Recombinant Rabbit Monoclonal
Anti-L-Lactyl-Histone H4 (Lys12) Rabbit mAb
Catalog Number: PTM-1411RM
$ 435

Clone Number: 37H4L1

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H4K12la

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 H4K12la
UniProt ID

P62805

Immunogen L-lactylated human histone H4 (Lys12) peptide
MW (kDa) 11
Specificity Anti-L-Lactyl-Histone H4 (Lys12) Rabbit mAb detects histone H4 when it is L-lactylated at Lys12.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
IP 1:50 - 1:100 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: 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: 10 seconds
The list of peptides used in the experiment is provided below.
Lane 1: H4K12 L-lactyl. Lane 2: H4K12 crotonyl.
Dot 3: H4K5 crotonyl. Dot 4: H4K5 2-hydroxyisobutyryl.
Dot 5: H4K5 L-lactyl. Dot 6: H4K8 crotonyl.
Dot 7: H4K8 L-lactyl. Dot 8: H4K8 2-hydroxyisobutyryl.
Dot 9: H4K12 2-hydroxyisobutyryl. Dot 10: H4K16 L-lactyl.
Dot 11: H4K12 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: 30 seconds
Predicted band size: 11 kDa
Observed band size: 11 kDa

Lysates: HeLa, NIH-3T3, BRL, Mouse liver
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: 11 kDa
Observed band size: 11 kDa
IHC-P

Tissue: Human testis
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: 1:1000
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-1411RM.

Tissue: Mouse liver
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: 1:1000
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-1411RM.

Tissue: Rat cerebrum
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: 1:1000
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-1411RM.
IP

IP of HeLa+ sodium lactate (100 mM, 24 hours) extracts
Lane 1: 5% Input
Lane 2: IP with PTM-1411RM
Lane 3: IP with Rabbit mAb IgG Isotype Control
IP Ab incubation condition: 4°C overnight, 1:100 dilution
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB Primary Ab incubation condition: PTM-1411RM, 2 hours at room temperature, 1:500 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP Conjugate)
Exposure time: 30 seconds
Predicted band size: 11 kDa
Observed band size: 11 kDa
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 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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References (5)
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  • BRAFV600E restructures cellular lactylation to promote anaplastic thyroid cancer proliferation
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  • Lactate Activates Germline and Cleavage Embryo Genes in Mouse Embryonic Stem Cells
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    Journal Cells
    Authors Qing Tian, et al.
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  • Overexpression of Tfap2a in Mouse Oocytes Impaired Spindle and Chromosome Organization
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    Journal INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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