EPO mRNA - BioReagent,Integrity of mRNA ≥85% (CE);1.0 mg/mL

Cat. No.: E1506464

GRADE & PURITY BioReagent, Integrity of mRNA ≥85% (CE);1.0 mg/mL

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Overview

Storage buffer: 1 mM Sodium citrate, pH 6.5

EPO mRNA encodes human erythropoietin (EPO) protein, a hormone that regulates red blood cell production. EPO also plays a role in wound healing and the brain's response to nerve injury. Karikó et al. showed that after in vivo transfection of EPO mRNA, reticulocyte count and hematocrit increased significantly. As a model drug, EPO mRNA can be used to study the design and optimization strategies of mRNA drugs. 

This product mimics mature mRNA with a 5'Cap 1 structure and a 3'poly(A) tail, and can be efficiently expressed both in vivo and in vitro.

Product Attributes

Product Concentration:	1mg/mL
Storage Solution:	1 mM Sodium Citrate, pH6.5
Full Length of mRNA Sequence:	859nt
Integrity:	≥85%
Product Purity:	Single mRNA Electrophoresis Band
Transfection Efficiency:	293T Cell Transfection

Transportation and Storage

This product is transported with dry ice and can be stored at -20℃~-80℃ for two years.

Precautions

• It can be dissolved at room temperature. After dissolution, it should be stored in an ice box or on an ice bath. During use, be careful to prevent RNase contamination and degradation. After use, it should be immediately stored at  -20℃~-80℃.
• This product is for scientific research use only, and shall not be used for clinical diagnosis or treatment, nor for food or drugs.
• For your safety and health, please wear a lab coat and disposable gloves during operation.

Specifications

Specifications & Purity

BioReagent,Integrity of mRNA ≥85% (CE);1.0 mg/mL

Stability And Storage

Store at -20℃~-80℃ long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent

Product Line

biological_products

Application Protocols

Item-specific validated protocols are not provided. Below are general/literature protocols for mRNA handling and expression; adjust per cell type/system.

A. mRNA transfection (lipid-based, 24‑well example):

• Thaw an aliquot on ice. Dilute desired mass (e.g., 100–500 ng) in 50 µL RNase‑free buffer.
• In a separate tube, dilute lipid reagent per vendor guidance in 50 µL buffer.
• Combine, mix gently, and incubate 10–15 min at room temperature to form complexes.
• Add dropwise to cells at 60–80% confluence in 0.5–1 mL complete medium.
• Incubate 24–48 h at 37 °C. Assay secreted EPO by ELISA; collect supernatant at defined timepoints. Include carrier‑only and no‑RNA controls.

B. Electroporation (suspension cells):

• Resuspend 1×10^6 cells in electroporation buffer with 0.5–2 µg mRNA.
• Pulse with program optimized for the cell type. Recover cells immediately into pre‑warmed media; incubate 12–48 h before analysis.

C. Integrity check (denaturing agarose):

• Mix 200–500 ng mRNA with denaturing gel loading dye.
• Run on 1–1.2% agarose with formaldehyde or glyoxal system; visualize under UV after safe nucleic acid stain.

Notes:

• Use RNase‑free plastics, tips, and buffers; wear fresh gloves.
• Aliquot stock to avoid repeat freeze–thaw (per product data).
• Item-specific concentration, buffer, capping/tailing, and modifications are Not specified; refer to CoA/Spec Sheet.

Biological Roles

This product encodes erythropoietin (EPO); notes below summarize EPO biology (literature). Item-specific sequence/UTR/modification details are Not specified for this item; refer to CoA/Spec Sheet.

• Function: EPO is a secreted glycoprotein hormone that regulates erythropoiesis. Binding to the erythropoietin receptor (EPOR) activates JAK2/STAT5 signaling, influencing survival, proliferation, and differentiation of erythroid progenitors.
• Expression: Predominantly produced by renal interstitial fibroblast‑like cells in adults; hypoxia stabilizes HIFs, upregulating EPO transcription.
• Structure: Mature human EPO is a ~30–34 kDa glycoprotein (165 aa) with multiple N‑linked glycosylation sites; glycan composition modulates serum half‑life and bioactivity (literature).
• Broader roles: Beyond erythropoiesis, EPOR expression has been reported in select non‑hematopoietic tissues, mediating context‑dependent signaling (literature observations; experimental systems vary).
• Research implications of mRNA delivery: mRNA enables transient EPO expression to study secretion, glycosylation processing, receptor engagement, and dose kinetics without genomic integration.

Note: The above describes endogenous biology of EPO. This SKU is a research reagent mRNA; it is not intended for clinical applications. Any item‑specific coding sequence (e.g., signal peptide, tags), codon optimization, or chemical base/sugar modifications are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

Applicability: Relevant. mRNA stability depends strongly on buffer selection and nuclease control.

General/literature guidance:

• Working solvents: RNase‑free water for short‑term use; for added stability, 10 mM Tris‑HCl, pH 7.2–8.0, with 0.1–1 mM EDTA to chelate Mg2+/Ca2+ that accelerate RNA hydrolysis. Item-specific buffer composition is Not specified for this item; refer to CoA/Spec Sheet.
• Salt effects: High ionic strength can promote structure/aggregation; keep NaCl ≤50–150 mM unless required by formulation, then assess stability.
• pH range: Keep near neutral (pH 7.0–8.0). Acidic conditions enhance depurination; alkaline conditions accelerate 2′‑OH transesterification.
• Additives: RNase inhibitors during manipulation; sugars (sucrose/trehalose 5–10%) can improve stability during freezing or lyophilization (general).

Example buffer recipes (literature; prepare RNase‑free):

• TE‑Low: 10 mM Tris‑HCl, 0.1 mM EDTA, pH 7.5.
• Citrate: 10 mM sodium citrate, pH 6.5–6.8 (commonly used in LNP formulation steps; exchange post‑encapsulation).
• Storage: Aliquot in RNase‑free water or TE‑Low; avoid repeated freeze–thaw (per product data).

Electrophoresis and QC:

• Run on denaturing agarose (e.g., glyoxal or formaldehyde) or capillary electrophoresis to assess integrity. Use nuclease‑free TAE/TBE equivalents when applicable.

Green Alternatives

Context: As an aqueous biopolymer reagent, EPO mRNA does not involve organic solvent use during routine handling. “Green alternatives” considerations focus on delivery modality and buffer systems rather than solvent swaps.

Modalities and trade‑offs (general/literature):

• Direct mRNA delivery vs. plasmid DNA:
  • mRNA avoids antibiotic selection and bacterial propagation (reduced waste/bioburden) and enables transient, transcription‑independent expression. However, cold‑chain intensity is higher and stability is lower than plasmid DNA.
• LNP systems:
  • Favor ethanol‑reduced microfluidic processes and recover ethanol post‑mixing to minimize VOC release. Choose ionizable lipids with improved biodegradability when possible.
• Buffers and plastics:
  • Use minimal single‑use plastics; adopt certified RNase‑free, recyclable consumables where available. Prepare RNase‑free buffers in bulk to reduce packaging.

Brief comparison (general):

• mRNA reagent in RNase‑free water: Aqueous; minimal solvent footprint; requires cold chain.
• Plasmid DNA alternative: Ambient‑stable options exist; requires transfection to nucleus and transcription; potential for antibiotic marker waste.
• Protein standard (purified EPO): No nucleic acid handling; but upstream expression/purification may entail larger solvent use.

Selection should balance environmental impact, experimental requirements (speed, transient kinetics), and infrastructure (cold storage). Item-specific green attributes are Not specified for this item; refer to CoA/Spec Sheet.

Pharmaceutical Uses

Clinical/therapeutic use is not claimed. For research use only (per product data).

Formulation and development research contexts (general/literature):

• Process development: Screening and optimizing LNP or polymeric nanoparticles for mRNA encapsulation, using EPO mRNA as a model payload to assess encapsulation efficiency, particle size/PDI, zeta potential, and potency readouts.
• Excipient compatibility studies: Evaluating buffers (citrate, acetate, Tris), sugars (sucrose, trehalose), amino acids (histidine, arginine), and chelators (EDTA) for mRNA stability during freezing, thawing, and agitation.
• Analytical method development: Establishing assays for mRNA integrity (CE, LC‑UV, Bioanalyzer), capping/tailed fraction, dsRNA content (ELISA), and residuals (solvents, lipids) post‑formulation.
• Device/interface studies: Jetting/atomization shear tests and filter compatibility for single‑use systems.

Regulatory note:

• This SKU is not GMP‑manufactured and is not qualified for human or veterinary use. No pharmacopeial monograph is specified. Any claims of sterility, endotoxin limits, or residual solvent levels are Not specified for this item; refer to CoA/Spec Sheet.

Use these data strictly for nonclinical, pre‑formulation, and analytical development experiments in controlled laboratory settings.

Physical Properties

Item-specific physicochemical specifications are not provided. For exact values, see: Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties for mRNA reagents:

• Physical state: Aqueous solution or lyophilized solid (common formats; item-specific format not specified).
• Solubility: Freely soluble in RNase-free water and low-ionic-strength, neutral pH buffers (e.g., 10 mM Tris, pH 7.2–8.0) (literature).
• pKa: Not applicable to the polymer as a whole; phosphate diester groups are polyanionic at neutral pH (literature).
• LogP: Not meaningful for polyanionic biopolymers.
• UV absorbance: Strong at 260 nm; A260 of 1.0 corresponds to ~40 µg/mL single-stranded RNA (literature). Item-specific extinction coefficient depends on length and composition and is Not specified for this item; refer to CoA/Spec Sheet.
• Thermal behavior: No defined melting point; thermal denaturation of secondary structure occurs over a broad range; prolonged exposure >65 °C leads to hydrolysis (literature).
• Density/refractive index/boiling point: Not applicable in practical terms for dilute aqueous solutions.

Sequence length and mass (literature context):

• Human EPO mature protein is 165 aa → coding region ~498 nt; with UTRs and poly(A), typical research-grade EPO mRNA constructs often range ~0.9–2.0 kb, giving masses roughly 0.3–0.7 MDa. Exact length/mass for this item: Not specified for this item; refer to CoA/Spec Sheet.

Quality and Grades

Item-specific quality designation: BioReagent (per product data).

What BioReagent grade implies (general context):

• Produced and packaged for research use in biological systems, emphasizing low bioburden, low endotoxin/pyrogen load, and nuclease control during manufacture and filling (typical of BioReagent-grade nucleic acids; exact acceptance criteria are manufacturer-specific and Not specified for this item; refer to CoA/Spec Sheet).
• Suitable for common molecular biology workflows (e.g., cell transfection, in vitro translation, RT‑qPCR standards), contingent on item-specific performance data, which are Not specified for this item; refer to CoA/Spec Sheet.

What is NOT claimed in absence of data:

• Clinical/diagnostic grade, GMP compliance, sterile apyrogenic certification, or specific capping/poly(A) configurations are not stated. Any stabilizers, sequence verification method, capping efficiency, tail length, or HPLC purity are Not specified for this item; refer to CoA/Spec Sheet.

Practical considerations:

• If UV trace, RIN value, or gel QC images are required, request the CoA for this lot.
• When comparing BioReagent-grade mRNA lots, evaluate: integrity (RIN/agarose), residual DNA/template carryover, dsRNA content, endotoxin (EU/µg), capping/tailing metrics, salt profile, and concentration. These parameters for this specific SKU are Not specified; refer to CoA/Spec Sheet.

Reaction and Applications

Manufacturer application notes: Not provided. The following are general/literature applications for EPO mRNA in research settings; item-specific performance is Not specified for this item; refer to CoA/Spec Sheet.

• Transient expression: Direct cytosolic delivery (e.g., lipid nanoparticles, cationic lipids, electroporation) to produce EPO protein without nuclear entry or transcription. Useful for studying EPO secretion pathways, glycosylation, and dose‑response kinetics.
• Positive control/standard: Calibration of RT‑qPCR assays targeting EPO mRNA sequences; spike‑in controls for RNA extraction/reverse transcription workflows.
• In vitro translation: Template for cell‑free systems (e.g., rabbit reticulocyte lysate, wheat germ) to assess translation efficiency and 5′/3′ UTR design variables (cap, Kozak, tail; specific features for this item Not specified).
• Protein engineering: Comparative studies of EPO variants (signal peptide optimization, codon optimization) using mRNA as the expression substrate (general approach).
• Formulation/process development: Screening lipid nanoparticle (LNP) or polymer carriers; assessing encapsulation efficiency, particle size, polydispersity, and in vitro potency readouts.

Practical tips (literature):

• Maintain RNase‑free technique; keep on ice during setup. Aliquot to avoid freeze–thaw.
• For cellular delivery, optimize dose (ng–µg per well), carrier ratio (N/P or w/w), and exposure time. Include no‑RNA and carrier‑only controls.
• Verify expression by ELISA for secreted EPO, western blot, or bioassays; confirm mRNA integrity by denaturing agarose or capillary electrophoresis.

Reaction Conditions

Item-specific tested conditions are not provided. The following are general/literature conditions for common mRNA workflows; optimize per system.

• Cell transfection (lipid-based):

  • Dose: 10–1000 ng mRNA per 24‑well (0.5–1 mL) culture; scale with surface area and cell type.
  • Carrier ratio: Follow lipid manufacturer’s N/P or µL reagent per µg mRNA guidelines; typical incubation 10–20 min for complexation.
  • Conditions: 37 °C, 5% CO2; media change after 3–6 h if cytotoxicity observed.
  • Readouts: Secreted EPO by ELISA at 12–72 h; cell viability controls included.

• Electroporation:

  • Dose: 0.1–10 µg per million cells depending on cell type.
  • Parameters: Program‑specific pulse settings; use manufacturer cuvettes/buffers; immediate transfer to pre‑warmed media.

• In vitro translation:

  • Systems: Rabbit reticulocyte or wheat germ extracts.
  • Input: 10–200 ng per 25 µL reaction; Mg2+ and K+ concentrations per kit; incubate 30–120 min at 25–30 °C (wheat germ) or 30 °C (reticulocyte).

• RT‑qPCR standards:

  • Prepare serial dilutions in RNase‑free carrier (e.g., tRNA) to minimize adsorption; reverse transcribe with random hexamers or gene‑specific primers.

Always maintain RNase‑free conditions, keep solutions on ice during setup, and avoid repeated freeze–thaw cycles (per product data). Exact buffer, concentration, and sequence for this SKU are Not specified; refer to CoA/Spec Sheet.

Safety and Handling

GHS classification, signal word, and hazard statements: Not specified for this item; refer to SDS for authoritative safety information.

General laboratory safety for mRNA reagents:

• Primary hazards: Low intrinsic toxicity; main risk is RNase contamination leading to degradation. Avoid aerosol generation when handling concentrated stocks.
• PPE: Laboratory coat, nitrile gloves (change frequently to avoid RNase transfer), and safety glasses. Use dedicated RNase-free consumables.
• Engineering controls: Work in a clean area; for sensitive applications, use a designated RNase-free biosafety cabinet. Treat work surfaces and tools with RNase decontamination solutions.
• Incompatibilities: RNases (ubiquitous on skin and common lab plastics), acidic/alkaline conditions (acid/base-catalyzed hydrolysis), divalent cations (can catalyze hydrolysis), and elevated temperatures. Avoid repeated freeze–thaw cycles (provided in product data).
• First aid (general): If contact with eyes/skin occurs, rinse with water. If inhaled as aerosol, move to fresh air. If ingested, rinse mouth with water. Seek medical attention if symptoms persist. Always consult SDS.
• Waste: Dispose according to institutional biosafety procedures for non-hazardous biologicals. Decontaminate surfaces with RNase-inactivating agents.

Shipping/storage risks:

• Shipped cold (ice chest + ice pads, per product data). Prolonged warm exposure degrades RNA; promptly store at the recommended temperature on receipt.

Solvent Selection

Applicability: mRNA is handled in aqueous, RNase‑free media; organic solvent selection is generally not applicable.

General guidance (literature):

• Preferred solvents/buffers: RNase‑free water; low‑salt, near‑neutral buffers (e.g., 10 mM Tris‑HCl, pH 7.2–8.0). Include 0.1–1 mM EDTA to chelate divalent cations that catalyze RNA hydrolysis. Magnesium is avoided unless required for downstream reactions.
• Avoid: Strong acid/base, high ionic strength without stabilizers, and most organic solvents (precipitation/denaturation). Short exposure to 70% ethanol/NaOAc is used only for precipitation/purification, not storage.
• Additives (use-case dependent): RNase inhibitors for handling; carrier RNA or glycogen during precipitation; sucrose/trehalose for lyophilization (item-specific stabilizers Not specified for this item; refer to CoA/Spec Sheet).

Comparison (general):

• RNase-free water: Maximal compatibility; lowest background for downstream enzymology.
• Tris-EDTA (TE): Improved chemical stability via chelation; may inhibit Mg2+-dependent enzymes; dilute before reactions requiring Mg2+.
• Citrate buffer pH 6.5–7.0: Sometimes used for LNP formulation to promote encapsulation; exchange post-formulation.

Choose solvents based on downstream application (transfection, RT‑qPCR, IVT/translation), while keeping ionic strength modest and pH near neutral.

Storage and Reconstitution

From product data:

• Storage: Store at −20 °C.
• Handling: Avoid repeated freezing and thawing.
• Shipping: Ice chest + ice pads.

General/literature best practices for mRNA reagents:

• Upon receipt: Inspect cold chain, immediately place at −20 °C (short‑term) or −80 °C (long‑term, preferred when available). Item-specific stability claims are Not specified for this item; refer to CoA/Spec Sheet.
• Aliquoting: Thaw once on ice, mix gently, spin briefly, and dispense into single‑use RNase‑free aliquots sized for one experiment (e.g., 5–20 µL) to prevent freeze–thaw cycles.
• Reconstitution: If lyophilized, reconstitute with RNase‑free water or low‑ionic TE‑Low (10 mM Tris‑HCl, 0.1 mM EDTA, pH 7.5). Target concentrations commonly range 0.1–1 mg/mL; item-specific recommended concentration/volume is Not specified for this item; refer to CoA/Spec Sheet.
• Storage buffer: Neutral pH, low ionic strength; optional 0.1–1 mM EDTA. Avoid divalent cations that accelerate hydrolysis.
• Working on ice: Keep solutions on ice during setup; minimize time at room temperature.
• Adsorption losses: Use low‑bind tubes and add carrier RNA for very dilute solutions.
• Freeze conditions: Snap‑freeze aliquots on dry ice/ethanol bath; store upright to reduce cap leakage.

Always consult the SDS and lot‑specific CoA for exact composition, concentration, and stability of SKU E1506464.

Structure and Identity

Item-specific data provided: minimal. Where exact values are unknown, see: Not specified for this item; refer to CoA/Spec Sheet.

• Product type: mRNA reagent encoding human erythropoietin (EPO)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet. (Varies with length and modification.)
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. (Scales with nucleotide count and any cap/tail/modifications.)
• SMILES: Not applicable for large heteropolymeric RNA.
• InChIKey: Not applicable for large heteropolymeric RNA.
• CAS/CID: Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general/literature):

• Linear, single‑stranded ribonucleic acid composed of A, C, G, U ribonucleotides linked by 3′→5′ phosphodiester bonds.
• 5′ region may be engineered (e.g., cap structures, UTRs); 3′ region may include a poly(A) tail and 3′ UTR elements (literature). Item-specific modifications: Not specified for this item; refer to CoA/Spec Sheet.
• Coding sequence: Encodes prepro‑EPO; the mature EPO protein is 165 aa (literature). The open reading frame is ~498 nt; total mRNA length depends on chosen UTRs and poly(A) tail (literature).

2D description in words (general):

• A continuous polymer backbone of alternating ribose and phosphate groups with pendant nucleobases (A/C/G/U). No defined small‑molecule ring system; stereochemistry fixed at the ribose centers (D‑ribose). Sequence order dictates the encoded EPO protein. Any cap/poly(A)/chemical modifications, if present, are located at the termini or within bases/sugar but are not specified for this item.

Synthetic Utility

Traditional small‑molecule synthetic utility does not apply. In molecular biology and bioengineering, EPO mRNA is a versatile substrate (general/literature):

• Protein production without transcription: Enables direct translation in cells or cell‑free systems, bypassing promoter dependency and nuclear transport.
• Reverse transcription and cloning: Serves as template for cDNA synthesis (RT) followed by PCR/cloning to analyze codon/UTR designs or to verify integrity (when sequence is known; item-specific sequence Not specified).
• UTR and modification studies: Systematic variation of 5′/3′ UTRs, cap structures, nucleotide chemistries (e.g., m1Ψ, 5‑moU), and poly(A) length to dissect structure–function relationships (general; not item-specific claims).
• Secretion pathway analysis: EPO carries a signal peptide; mRNA transfection supports studies on signal peptide efficiency, ER targeting, glycosylation, and secretion rates.
• Reporter fusions: In research, EPO coding sequence can be fused to tags or reporters to quantify expression/secretion; any such features for this item are Not specified for this item; refer to CoA/Spec Sheet.

These utilities make EPO mRNA a robust tool for expression benchmarking and formulation screening where a secreted, quantifiable protein readout is advantageous.

Target Specificity

Not applicable. This SKU is an mRNA reagent and not an antibody, probe, or assay kit with defined antigen/epitope specificity. No clone, isotype, or species reactivity information applies. For sequence-level details (e.g., species variant of EPO, codon optimization, UTRs), item-specific information is Not specified for this item; refer to CoA/Spec Sheet.

Frequently Asked Questions

What does BioReagent mean?

BioReagent indicates that the product has been processed and tested for biological and biochemical applications. Specifications may include endotoxin limits, microbial controls, sterility, low DNase/RNase/protease activity, and verified biological activity where relevant.

How should this product be stored?

Store at ?20 °C. Avoid repeated freeze–thaw cycles. Aliquot into single-use volumes before freezing.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.