Creative Biolabs offers an expansive suite of pan-acetylation and site-specific acetylation antibodies using proprietary High-Affi™ technology, serving as critical tools for identifying, verifying, and quantifying protein acetylation events.

Background

Protein acetylation ranks among the most frequent post-translational modifications (PTMs) observed in biological systems. This biochemical reaction transfers acetyl groups (CH3C=O, structurally abbreviated as Ac) sourced from acetyl-coenzyme A to either N-terminal residues or lysine side-chain ε-amino groups.

N-terminal acetylation is an evolutionarily conserved modification found in nearly all eukaryotic proteins and regulatory peptides, as well as certain prokaryotic proteins. Multiple N-terminal acetyltransferases (NATs) drive this typically permanent modification. Despite affecting 80 to 90 percent of proteins and engaging in documented molecular mechanisms, its comprehensive biological significance remains partially unresolved. Established findings reveal effects of N-terminal acetylation on protein structural organization, intracellular transport pathways, molecular interaction networks, and diverse cellular operations.

Fig.1 Schematic outline of N-terminal acetylation of proteins.¹

Lysine acetylation is a reversible, precisely controlled modification occurring at numerous sites across heterogeneous proteins. Histone lysine acetylation (H2A, H2B, H3, H4) directly enables chromatin unwinding and governs architectural configuration and functional adaptability. Non-histone targets encompass transcription regulators, chaperone proteins, signaling molecules, and structural components. Opposing enzymatic activities of lysine acetyltransferases (KATs) and deacetylases (KDACs) modulate this modification, affecting metabolic control, energy management, and genetic regulation. Cellular stress conditions markedly elevate p53 acetylation, activating tumor-suppressive functions via growth inhibition. Disrupted acetylation patterns link to pathologies such as cancers, metabolic syndromes, neurodegenerative diseases, and immune disorders, highlighting therapeutic potential through acetylation modulation.

Fig.2 Schematic outline of lysine protein acetylation.¹

Antibody Types

Based on Clonality:

Monoclonal Antibodies: Generated through clonal expansion of single immune cells, these agents target individual molecular signatures (specific binding regions on acetylated proteins). Their uniformity across production batches ensures precise experimental reproducibility.

Polyclonal Antibodies: Harvested from multiple clones of immune cells, these contain heterogeneous antibody populations recognizing multiple antigenic determinants on the acetylated protein. While cost-effective for large-scale production, inherent biological variability may introduce inter-batch signal fluctuations and nonspecific interactions.

Recombinant Antibodies: Constructed through DNA manipulation in host systems, these combine genetic engineering precision with manufacturing scalability. They offer advantages in terms of reproducibility, scalability, and potential for engineering specific properties.

Based on Specificity:

Pan-Acetylation Antibodies: Identify acetylated lysine moieties independent of protein context or adjacent sequences, serving as screening tools for global acetylation pattern shifts.

Site-Specific Acetylation Antibodies: Engineered to bind acetylation events at predetermined lysine locations within defined protein architectures. Critical for mechanistic studies interrogating site-specific modification impacts.

Discovery Strategy

Creative Biolabs engineers high-precision acetylation-targeting antibodies through integrated phage display systems and synthetic peptide immunization protocols, utilizing lysine-containing acetylated peptide constructs.

Monospecific Anti-Acetylation Polyclonal Antibody Production

Creating monospecific polyclonal antibodies initiates with animal immunization using precisely engineered immunogens—acetylated peptides coupled to carrier proteins. Host immune systems generate diverse antibody populations, with subsets recognizing acetylated epitopes. Subsequent purification employs sequential affinity chromatography: initial exposure to non-acetylated peptide columns removes cross-reactive antibodies, followed by acetylated peptide column retention of target-specific immunoglobulins. Elution yields antibody preparations enriched for acetylation specificity.

Phage Display Strategy for Anti-Acetylation Monoclonal Antibody Discovery

This in vitro selection technique leverages bacteriophage libraries displaying variable antibody fragments (scFv/Fab domains). Acetylation-focused biopanning cycles commence with phage incubation on immobilized acetylated peptides. Non-adherent phages undergo elimination, while binders undergo amplification. Intermediate counter-screening with non-acetylated variants enhances specificity. Post multiple selection rounds, isolated phage clones undergo DNA sequencing to identify antibody-coding sequences, enabling recombinant production of acetylation-specific monoclonals.

Hybridoma Strategy for Anti-Acetylation Monoclonal Antibody Discovery

The established hybridoma protocol initiates with customized acetylated antigen immunization in murine hosts. Post-immune response maturation, splenic B lymphocytes fuse with immortalized myeloma counterparts, generating hybrid cell lines. These hybridomas retain antibody secretion capacity while gaining perpetual proliferative potential. High-throughput screening isolates clones producing acetylation-specific antibodies, followed by monoclonality verification through subcloning. Validated lines establish continuous antibody production systems.

Types of PTM

Beyond acetylation, we facilitate custom antibody development for diverse PTMs.

Service Highlights

Epitope Precision: Strategic immunogen design and multi-phase screening minimize cross-reactivity, guaranteeing modification-specific detection.

Dual Development Systems: Polyclonal and phage display platforms address varied research needs balancing signal intensity and reproducibility.

Adaptable Workflows: Collaborative planning from immunogen conception to bulk production.

Multilayer Validation: Specificity verification via ELISA, Western blot, and comparative peptide/protein analyses.

Technical Partnership: Continuous scientific support throughout development cycles.

Cases

Monospecific Anti-Acetylation Polyclonal Antibody Production

• QC analysis certificate of the synthesized acetylated peptide

• Specificity test of anti-acetylated peptide polyclonal antibody by Dot blot

Q&A

Q: How are acetylation-specific antibodies used in research?

A: These specialized antibodies serve as essential investigative instruments across biological disciplines. Their capacity to identify and measure protein acetylation events supports mechanistic studies of cellular communication networks, genetic control systems, chromatin reorganization processes, and enzymatic functionality. Scientists apply these tools in Western analysis, fluorescence-based imaging, chromatin isolation techniques (ChIP), and large-scale protein identification methods to examine acetylation's changing roles in health conditions like malignancies, neural degradation, and immune responses.

Q: How would you define anti-lysine acetylation antibodies?

A: These immunological reagents specifically detect lysine residues modified by acetylation - a biochemical alteration where proteins gain acetyl groups (CH₃CO−) at their ε-amino positions. Engineered through stringent development protocols, they discriminate between acetylated lysines and other molecular variants, becoming vital for mapping acetylation distribution and functional impacts within living systems.

Q: What quality controls guarantee antibody specificity?

A: Our multi-tiered validation framework begins with strategic immunogen construction, integrating target acetyl sequences while minimizing structural similarities to unrelated proteins. Screening phases employ comparative analyses using modified and unmodified peptides through ELISA and blotting techniques. Final verification stages test antibody performance across experimental models, including treated versus untreated cellular samples, ensuring exclusive recognition of intended targets.

Q: What's the expected development timeframe for custom antibodies?

A: Project durations reflect biological complexity and platform selection. Polyclonal development typically spans 8-16 weeks, whereas monoclonal generation through phage display or hybridoma techniques may require 16-32 weeks for initial characterization. We establish clear temporal expectations during project planning while maintaining regular progress updates throughout development cycles.

Q: Can you create antibodies targeting novel acetylation sites?

A: Our platform specializes in developing antibodies against researcher-identified acetylation positions. Clients submit target sequences containing modified lysines, which our team translates into optimized immunogenic constructs. With extensive experience across protein families, we deliver antibodies precisely aligned with experimental objectives, whether investigating established or newly discovered acetylation sites.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.