HNG peptide ([Gly14]-Humanin) is a highly potent synthetic derivative of the mitochondrial-derived micropeptide Humanin, modified via a single amino acid substitution (Ser14Gly). Supplied strictly as a research compound for in vitro and preclinical laboratory investigation, HNG exhibits up to 1,000-fold greater biological activity than its parent molecule in cytoprotective and anti-apoptotic assay models.
HNG peptide ([Gly14]-Humanin) is a highly potent synthetic derivative of the mitochondrial-derived micropeptide Humanin, modified via a single amino acid substitution (Ser14Gly). Supplied strictly as a research compound for in vitro and preclinical laboratory investigation, HNG exhibits up to 1,000-fold greater biological activity than its parent molecule in cytoprotective and anti-apoptotic assay models.
HNG peptide, chemically designated as [Gly14]-Humanin, is an engineered analogue of Humanin—a 24-amino acid micropeptide encoded within the 16S ribosomal RNA region of the mitochondrial genome. In native Humanin, serine occupies position 14 of the peptide sequence (MAPRGFSCLLLLTSEIDLPVKRRA). By replacing serine with glycine at this specific loci, researchers synthesized HNG (MAPRGFSCLLLLTGEIDLPVKRRA), resulting in a dramatic increase in biological potency across multiple cellular models.
As a fundamental subject in mitochondrial biology, the HNG peptide is classified as a mitochondrial-derived peptide (MDP). Native MDPs act as retrograde signaling molecules, communicating cellular stress states from the mitochondria to the nucleus and extracellular matrix. Synthetic modification to yield HNG preserves these endogenous signaling pathways while enhancing receptor binding affinity and peptide stability during extended in vitro culture conditions.
In academic and pharmaceutical research environments, HNG is utilized exclusively as a research compound for laboratory investigation. It serves as a tool for probing apoptosis pathways, cell survival cascades, oxidative stress responses, and mitochondrial membrane integrity without the confounding degradation profiles often observed with wild-type micropeptides.
Preclinical studies suggest that HNG mediates its potent cytoprotective effects through two primary cell-surface receptor configurations: the formyl peptide receptor-like 1 (FPRL1, also known as FPR2) and a heterotrimeric receptor complex comprising the ciliary neurotrophic factor receptor alpha (CNTFRα), interleukin-27 receptor subunit alpha (WSX-1), and glycoprotein 130 (gp130).
Upon ligand binding to the FPRL1/FPR2 receptor, HNG initiates intracellular signaling cascades that activate extracellular signal-regulated kinase (ERK1/2) and protein kinase B (Akt). Concurrently, engagement with the CNTFRα/WSX-1/gp130 complex triggers the Janus kinase 2 (JAK2) pathway, leading to the phosphorylation and nuclear translocation of Signal Transducer and Activator of Transcription 3 (STAT3). STAT3 activation induces the transcription of anti-apoptotic proteins, including Bcl-2 and Bcl-xL.
In vitro data indicate that HNG directly interferes with the intrinsic apoptotic pathway by neutralizing pro-apoptotic Bcl-2 family proteins. Specifically, HNG binds directly to Bax (Bcl-2-associated X protein), preventing its conformational change, oligomerization, and subsequent translocation to the outer mitochondrial membrane. By preventing Bax-mediated mitochondrial outer membrane permeabilization (MOMP), HNG inhibits the release of cytochrome c into the cytosol, thereby halting executioner caspase-3 and caspase-7 activation.
The scientific literature surrounding HNG spans diverse fields including neurobiology, cardiovascular pathology, metabolic disease, and cellular senescence. Because native Humanin demonstrated protective capacity against amyloid-beta peptide toxicity in original neuronal culture screens, HNG was rapidly evaluated across enhanced toxicity models.
In neurobiological in vitro models, preclinical data demonstrate that picomolar to nanomolar concentrations of HNG protect primary cortical neurons, hippocampal cultures, and PC12 cell lines against toxic insults induced by oligomeric amyloid-beta (Aβ1-42), N-methyl-D-aspartate (NMDA) excitotoxicity, and hypoxia-reoxygenation. Research indicates that HNG attenuates reactive oxygen species (ROS) accumulation and maintains mitochondrial membrane potential (ΔΨm) under severe oxidative stress.
In vascular and metabolic preclinical models, rodent studies suggest that HNG administration mitigates endothelial dysfunction and reduces atherosclerotic lesion progression. Investigations utilizing rodent models of myocardial infarction and ischemic stroke report that HNG treatment decreases infarct size, suppresses local inflammatory cytokine expression (such as TNF-α and IL-6), and preserves microvascular perfusion following reperfusion injury.
To contextualize HNG within the broader ecosystem of mitochondrial-targeted peptides, it is useful to compare its structural and functional parameters against related compounds evaluated in laboratory research.
When evaluated alongside wild-type Humanin, HNG demonstrates identical receptor targets but displays up to a 1,000-fold increase in protective efficacy in cell viability assays. Where native Humanin requires micromolar concentrations (1–10 µM) to elicit full anti-apoptotic protection, HNG achieves comparable cytoprotection at nanomolar or picomolar levels (1–10 nM).
Compared to other mitochondrial-derived signaling agents like MOTS-c, which primarily functions as a metabolic regulator influencing nuclear gene expression related to folate metabolism and glucose homeostasis, HNG acts primarily as a direct anti-apoptotic factor and receptor-mediated survival agent. Meanwhile, synthetic targeted compounds like SS-31 (Elamipretide) act directly within the inner mitochondrial membrane by binding cardiolipin to optimize electron transport chain kinetics, whereas HNG relies predominantly on membrane receptor engagement and signal transduction.
Researchers exploring comprehensive mitochondrial survival pathways often cross-reference these distinct mechanisms within the PX1 Research Library to select the appropriate compound for specific experimental end-points.
Proper reconstitution technique is critical to maintaining peptide solubility, secondary structure, and biological availability during laboratory experimentation. HNG is supplied as a lyophilized (freeze-dried) powder requiring strict aseptic handling.
Prior to reconstitution, the vial should be allowed to equilibrate to room temperature (18°C to 25°C) inside a laminar flow hood to minimize condensation formation upon opening. The recommended primary solvent for research-grade HNG is sterile, non-bacteriostatic water for injection or sterile phosphate-buffered saline (PBS, pH 7.4). For assays sensitive to trace salts, initial dissolution in sterile molecular-grade water followed by dilution into experimental culture media is advised.
Gently swirl or invert the vial to dissolve the lyophilized cake. Avoid violent vortexing or aggressive sonication, as mechanical shear stress can disrupt peptide secondary structure or promote aggregation. Reconstitute to a stock concentration (e.g., 1 mg/mL or 1 mM) that permits accurate volumetric pipetting into final culture assays while minimizing DMSO or vehicle carrier concentration in working media.
To preserve the structural integrity and binding capability of HNG over extended experimental timelines, strict temperature management and physical storage conditions must be maintained.
Lyophilized HNG peptide should be stored in a commercial freezer at -20°C for short-to-medium-term storage (up to 12 months) or at -80°C for long-term storage (up to 24 months). Store vials in desiccated containers away from light exposure to prevent photo-degradation and moisture absorption.
Once reconstituted into aqueous solution, stock solutions of HNG should be aliquoted into single-use microcentrifuge tubes (polypropylene, low-binding) to prevent repeated freeze-thaw cycles, which degrade peptide purity over time. Aliquoted aqueous solutions are stable at -80°C for up to 3 to 6 months. Diluted working solutions in tissue culture media should be prepared fresh daily and kept on ice during active laboratory procedures.
The validity of preclinical assay data depends directly on the chemical purity and structural identity of the research compounds utilized. Impurities such as truncated peptide sequences, residual TFA salts, or endotoxins can alter cell viability metrics, yield false-positive results, or induce non-specific inflammatory signaling in vitro.
PX1 Research enforces stringent quality control measures for all catalog compounds, including all research peptides. Every production lot of HNG undergoes rigorous analytical testing prior to release:
1. Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC): Establishes chemical purity, guaranteeing a minimum of 98.0% purity for every lot. HPLC chromatograms must display a sharp, single major peak with minimal secondary signal. 2. Electrospray Ionization Mass Spectrometry (ESI-MS): Confirms exact molecular weight (2,637.1 Da calculated) and verifies sequence accuracy without structural modifications or sequence truncations. 3. Chromogenic LAL Endotoxin Analysis: Quantifies bacterial endotoxin levels to ensure values remain well below standard cell culture thresholds (<0.01 EU/mg), preventing confounding toll-like receptor (TLR) activation in immunologically sensitive assays.
Every shipment from PX1 Research includes a lot-specific Certificate of Analysis (COA) directly linked to these analytical reports, ensuring full traceability from synthesis to laboratory receipt.
PX1 Research operates dedicated manufacturing and distribution facilities within the United States. Compounds are synthesized in GMP-compliant facilities utilizing automated solid-phase peptide synthesis (SPPS) platforms, followed by preparative HPLC purification and controlled freeze-drying protocols.
All analytical testing is executed in an ISO 17025 accredited testing laboratory, providing independent, unbiased verification of physical parameters, heavy metal content, and biological cleanliness. Vials are sealed under inert argon atmosphere to eliminate oxygen exposure prior to distribution.
Orders placed through PX1 Research are processed and shipped same-day (Monday through Friday) from centralized fulfillment nodes in California and Arizona. Fast, temperature-controlled transit prevents exposure to environmental extremes during transit. Institutional buyers and primary investigators seeking scaled batch quantities for multi-center research programs can utilize our bulk lab account program for custom synthesis lots and dedicated inventory reserves.
What is HNG peptide and how does it differ from native Humanin?
HNG peptide ([Gly14]-Humanin) is a synthetic derivative of the mitochondrial-derived peptide Humanin. It differs by a single amino acid substitution at position 14, where serine is replaced by glycine. In vitro assays demonstrate that HNG possesses up to 1,000-fold greater cytoprotective potency compared to wild-type Humanin.
What is the molecular weight and sequence of HNG?
HNG has a molecular sequence of MAPRGFSCLLLLTGEIDLPVKRRA and a theoretical molecular weight of approximately 2,637.1 Da. Identity is verified via ESI-MS or MALDI-TOF mass spectrometry.
Is HNG peptide approved for human clinical use or therapy?
No. HNG is supplied strictly as a research compound for in vitro laboratory experiments and preclinical animal research. It is not approved by the FDA or any regulatory body for human consumption, clinical treatment, diagnostic use, or therapeutic administration.
How should HNG peptide be reconstituted for cell culture assays?
Reconstitute lyophilized HNG in sterile, non-bacteriostatic water or sterile PBS (pH 7.4). Allow the vial to warm to room temperature before opening, add the diluent slowly along the vial wall, and gently invert until fully dissolved. Avoid aggressive vortexing.
What are the recommended storage temperatures for HNG?
Lyophilized HNG powder should be stored at -20°C or -80°C in a dry, dark environment. Once reconstituted, stock aliquots should be stored at -80°C to prevent degradation and avoid multiple freeze-thaw cycles.
How does PX1 Research verify the purity of HNG peptide?
PX1 Research subjects every batch of HNG to RP-HPLC purity analysis (guaranteeing ≥98% purity), mass spectrometry for sequence identity, and LAL assays for endotoxin quantification (<0.01 EU/mg). Certificates of Analysis (COA) are provided for every lot.
What primary receptor pathways are activated by HNG in preclinical models?
In vitro studies indicate HNG interacts with the FPRL1/FPR2 cell-surface receptor and the CNTFRα/WSX-1/gp130 receptor complex, activating downstream ERK, Akt, and STAT3 signaling pathways to inhibit Bax-mediated apoptosis.
Can I obtain bulk or institutional quantities of HNG from PX1 Research?
Yes. Institutional researchers, university labs, and corporate R&D departments can request bulk synthesis or lot-reserved quantities through the PX1 Research wholesale portal.
All products are sold strictly for laboratory and research use only. Not for human or veterinary use, diagnosis, treatment or consumption. Statements have not been evaluated by the FDA.