Hemoglobin Solution (10 mg/mL) - BioReagent

Cat. No.: H1528851

GRADE & PURITY BioReagent

Storage

Store at 2-8°C

Shipped In

Wet ice

Overview

This product is designed as a positive control for the qualitative detection of fecal occult blood. The formulation consists primarily of bovine hemoglobin, deionized water, and a preservative system. This reagent is supplied for research purposes exclusively and is not indicated for clinical diagnostic procedures or any other applications.

Specifications

Specifications & Purity

BioReagent

Storage

Store at 2-8°C

Shipped In

Wet ice

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

Grade

BioReagent

Product Line

Biological Products

Application Protocols

Below are example research protocols leveraging hemoglobin’s optical properties and redox chemistry. These are general literature-style guidelines; optimize for your system.

1. Quick spectral check of state (oxy vs. met):

• Dilute an aliquot 1:50–1:200 in air-equilibrated phosphate buffer (pH ~7.4) in a quartz cuvette.
• Record 350–700 nm spectrum. OxyHb shows Soret ~415 nm with α/β bands near 577/541 nm; metHb Soret ~405 nm and altered visible bands.
• If met content is high, reduce gently (e.g., small excess dithionite), then re-equilibrate with air for oxyHb and re-measure.

2. Preparation of deoxyhemoglobin for kinetics:

• Degas buffer thoroughly (freeze–pump–thaw or argon sparge). Keep all solutions on ice.
• Add minimal dithionite to oxyHb to form deoxyHb; verify by Soret shift (~430 nm) and visible spectrum.
• Transfer to an anaerobic, gas-tight cuvette or stopped-flow syringe for ligand-binding measurements.

3. Pseudoperoxidase assay (initial-rate):

• In phosphate buffer pH 7.0 at 25°C, mix hemoglobin (e.g., 0.1–1 µM heme sites) with chromogenic substrate (e.g., 1–5 mM guaiacol) and initiate with 50–200 µM H2O2.
• Monitor product formation at the appropriate wavelength (e.g., 470 nm for tetraguaiacol). Stop reaction before significant heme bleaching.

Note: Concentration, buffer composition, and preservatives for this item are Not specified; verify compatibility and adjust dilutions accordingly. Research use only.

Biological Roles

General biological context (literature):

• Function: Hemoglobin is the principal oxygen transport protein in vertebrate erythrocytes, binding O2 at ferrous heme iron and releasing it in tissues. It exhibits cooperative binding and allosteric regulation by protons (Bohr effect), CO2 (carbamino formation), and organic phosphates (e.g., 2,3-BPG in many mammals).
• Structure–function: The α2β2 tetramer can adopt tense (T) and relaxed (R) conformations, with ligand binding shifting the equilibrium toward R, enhancing affinity at remaining sites. Heme’s electronic transitions underpin the characteristic UV–Vis spectra used for state assignment.
• Redox states: Fe(II) form binds O2, CO, and NO; oxidation to Fe(III) yields methemoglobin, which cannot bind O2 but has distinct spectral and ligand properties. Reduction systems (e.g., cytochrome b5 reductase in vivo) maintain functional Fe(II).
• Gas signaling interactions: Hemoglobin can scavenge NO and bind CO; such interactions influence vascular signaling in physiology. In vitro, CO and NO ligation are widely used to probe heme pocket dynamics and ligand kinetics.
• Variants: Species- and isoform-specific differences in sequence and 2,3-BPG sensitivity modulate oxygen affinity; these differences should be considered when interpreting biophysical data.

Note: The biological origin, species, and isoform for this item are Not specified; refer to CoA/Spec Sheet if variant-specific properties are critical to your application.

Buffer Applications

This product is a protein solution and is not used as a buffering agent. However, it is commonly handled within standard biological buffers.

Practical notes (general):

• Compatible buffers: Phosphate, HEPES, or Tris at pH ~7.0–7.5 are typical for maintaining hemoglobin’s structural integrity and consistent spectral properties.
• Avoid: Strong chelators or denaturants (e.g., EDTA at high concentrations, urea, guanidinium) if native quaternary structure or heme environment must be preserved. High concentrations of imidazole or primary amines can interfere with certain spectroscopic/kinetic assays.
• Oxygen control: For deoxy or CO/NO studies, degas buffers thoroughly and use gas-tight vessels. For oxyhemoglobin, equilibrate with air or O2-saturated buffer.

If buffer formulation details of this item are required (salt, pH, preservatives), they are Not specified here; consult the CoA/Spec Sheet.

Green Alternatives

For an aqueous protein reagent, “green” considerations focus on formulation, shipping, and waste minimization rather than solvent replacement.

Greener practice options (general):

• Lyophilized format vs. solution:
  • Lyophilized hemoglobin (when available) can reduce cold-chain energy use and shipping mass; reconstitute on demand. Tradeoff: requires careful reconstitution to preserve functional state and may show greater lot-to-lot variability in oxidation state.
• Buffer selection:
  • Prefer phosphate or HEPES buffers over volatile amines in open systems to minimize air emissions; avoid heavy-metal-containing stabilizers. Keep ionic strength moderate to reduce salt waste.
• Additive minimization:
  • Use the lowest effective concentration of preservatives; consider single-use aliquots to avoid repeated freeze–thaw instead of adding biocides. Verify downstream assay compatibility.
• Waste management:
  • Aqueous protein waste can typically be deactivated with biodegradable detergents and disposed per local regulations. Avoid introducing unnecessary organic solvents which complicate disposal.

Comparison (general):

• Aqueous hemoglobin solution: Ready-to-use; requires refrigerated shipping/storage; potential preservative content (not specified for this item).
• Lyophilized hemoglobin: No cold shipping needed; longer shelf life at ambient prior to reconstitution; requires careful dissolution and oxygenation control to obtain desired redox/ligand state.

Item-specific formulation details (buffer, preservatives) are Not specified for this item; consult the CoA to tailor green handling choices.

Pharmaceutical Uses

No pharmaceutical or clinical use is claimed for this product. It is supplied strictly for research use only.

Context (general information):

• Proteins like hemoglobin are used in research related to formulation science, device calibration, and biophysical method development. However, excipient roles or pharmacopeial compliance for hemoglobin preparations are specialized and not implied here.
• This listing provides no information on GMP status, sterility, pyrogen/endotoxin levels, or compendial specifications.

Item-specific statement:

• Research Use Note: For research use only.
• Any application in manufacturing, diagnostic, or therapeutic contexts would require additional qualification not provided with this BioReagent-grade product.

Physical Properties

Item-specific physical specifications are not provided beyond concentration and storage; consult the CoA/SDS for definitive values.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration: 10 mg/mL (aqueous)
• Density, refractive index, viscosity: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Readily soluble in aqueous buffers; insoluble in nonpolar organic solvents (general, literature).
• pI (isoelectric point): Typically in the pH ~6.8–7.2 range for human HbA variants (literature; species/variant dependent).
• UV–Vis characteristics (literature):
  • Oxyhemoglobin exhibits a Soret band near ~415 nm with α/β bands near ~577/541 nm.
  • Deoxyhemoglobin Soret shifts to ~430 nm with diminished visible bands; methemoglobin Soret near ~405 nm.
• Approximate molecular mass (literature): ~64–65 kDa for tetrameric adult hemoglobin (species dependent; each subunit ~15–16 kDa plus heme).

Practical handling notes (general):

• Absorbance-based quantification often uses literature extinction coefficients (e.g., εSoret varies by redox/ligand state); verify state and buffer before calculation.
• Hemoglobin’s optical spectra are highly state-dependent (oxy-, deoxy-, met-, CO-bound); maintain controlled redox and gas atmosphere for reproducible measurements.

Quality & Grades

• Grade/Purity: BioReagent — suitable for general laboratory research applications, assay development, and biochemical studies. This designation typically indicates manufacturing and QC controls aligned with research use but not necessarily with clinical/diagnostic specifications.

What BioReagent commonly implies (general):

• Protein integrity and concentration are controlled within research-use limits; exact acceptance criteria vary by lot/manufacturer and are not specified here.
• Microbial/bioburden control may be implemented, but sterility is not guaranteed unless explicitly stated (not specified for this item).
• Low levels of interfering contaminants for routine biochemical work, but not trace-element/clinical grade certification.

Item-specific notes:

• Concentration: 10 mg/mL (aqueous). Buffer composition, preservative content (e.g., sodium azide), and exact hemoglobin source/variant are not provided — Not specified for this item; refer to CoA/Spec Sheet.
• UV cutoff, residual metals, protease activity, or endotoxin limits: Not specified for this item; refer to CoA/Spec Sheet.

Practical guidance:

• For quantitative spectroscopy or enzymology, confirm lot-specific extinction coefficients, redox/ligand state (oxy-, deoxy-, or met-), and buffer composition from the CoA.
• If using in cell-based assays or sensitive catalysis, verify presence/absence of preservatives and perform a quick spectral quality check (Soret/visible bands) upon receipt.

Reaction & Applications

Typical research applications of hemoglobin solutions leverage its heme-based ligand binding and redox chemistry (general literature guidance):

• Spectroscopic standards and calibration:
  • Generate oxy-, deoxy-, met-, and CO-bound forms to calibrate UV–Vis spectrometers (Soret and visible α/β bands) and pulse oximetry research devices. Maintain controlled gas atmospheres and verify state via characteristic spectra.
• Redox and peroxidase-like assays:
  • The heme center can catalyze peroxidative reactions in the presence of H2O2 (pseudoperoxidase activity). Use low H2O2 equivalents and monitor for heme degradation (spectral bleaching) over time.
• Nitric oxide/CO/O2 binding studies:
  • Quantify ligand association/dissociation kinetics with stopped-flow or equilibrium methods; buffer degassing and gas-tight cuvettes are recommended.
• Heme quantification and protein standards:
  • Serve as a positive control for heme assays or as a mass standard in heme–protein interaction studies.
• Chemical modification and crosslinking research:
  • Explore 2,3-BPG analog effects or site-selective modifications on cysteine/lysine residues (verify variant composition). Avoid over-oxidation which can impair tetramer stability.

Practical tips:

• Maintain cold chain (2–8°C) and minimize light exposure to reduce auto-oxidation to methemoglobin.
• Degas buffers for deoxy studies; add dithionite freshly and remove excess prior to certain measurements to avoid spectral interference.
• Record baseline spectra upon receipt to document lot-specific characteristics. Use low-bind plasticware to minimize loss at low concentrations.

Reaction Conditions

Because this is a protein reagent, “reaction conditions” pertain to biochemical/biophysical assays rather than classic synthetic transformations. General literature guidance is provided below; verify details empirically.

• Spectral state interconversion:
  • Oxy → deoxy: Degas buffer (pH ~7.0), add a slight excess of sodium dithionite under inert gas; remove excess reductant before certain measurements (e.g., via gel filtration) to avoid interference.
  • Deoxy → CO-bound: Bubble CO gently through deoxy solution at 0–4°C; monitor Soret/visible shifts to completion. Ensure proper ventilation and CO safety controls.
  • Fe(II) → metHb (Fe(III)): Treat with mild oxidants (e.g., ferricyanide) at neutral pH; remove oxidant; confirm by spectrum (~405 nm Soret).
• Pseudoperoxidase assays:
  • Typical conditions: pH 6.8–7.4 phosphate buffer, substrate (e.g., ABTS, guaiacol), and low millimolar H2O2. Start with sub-micromolar heme sites to minimize heme degradation; record initial rates at 25°C.
• Kinetics:
  • Use stopped-flow at 10–25°C for rapid CO/NO association rates; maintain anaerobic conditions and precisely control ligand concentrations.
• Stability window:
  • Temperature: Keep on ice or 2–8°C during setup; room temperature exposure should be minimized to reduce autoxidation.
  • Light: Limit strong light to reduce photodissociation and photo-oxidation during sensitive measurements.

Note: Buffer composition, preservatives, and exact variant for this item are Not specified; adjust conditions to your system and confirm spectral state before data collection.

Safety & Handling

Hazard data for this specific item are not listed here. Always consult the SDS for authoritative safety information.

• GHS classification: Not specified for this item; refer to SDS.
• Signal word / H-statements / Pictograms: Not specified for this item; refer to SDS.

General laboratory safety for protein aqueous solutions:

• PPE: Laboratory coat, safety glasses, and appropriate gloves (e.g., nitrile). Avoid inhalation of aerosols and contact with skin/eyes.
• Handling: Work using clean, protein-compatible plasticware or low-bind tubes to minimize adsorption. Avoid microbial contamination (use aseptic technique if sterility is required). Do not mouth-pipette.
• Incompatibilities: Strong oxidizers (promote methemoglobin formation), proteases (degradation), extreme pH (<5 or >9 can denature), and repeated freeze–thaw (aggregation/precipitation). Avoid azide-sensitive downstream applications if solution contains azide (not specified for this item).
• First aid (overview, general):
  • Eye/skin contact: Rinse with water. Remove contaminated clothing.
  • Ingestion/inhalation: Rinse mouth; move to fresh air. Seek medical attention if symptoms persist.
• Spill response: Wipe with absorbent material and clean with aqueous detergent; dispose of according to institutional biosafe/chemical waste protocols.
• Biological origin note: If derived from biological material, handle as a potential low-risk bio-macromolecule unless otherwise stated; do not assume sterility.

Storage per Product Data: Store at 2–8°C. Ship on wet ice.

Solvent Selection

This product is supplied as an aqueous protein solution (10 mg/mL). “Solvent selection” pertains mainly to compatible buffers and additives rather than organic solvents.

• Primary medium: Water-based buffers (e.g., phosphate, Tris, HEPES). Organic solvents are generally unsuitable and may denature hemoglobin.
• pH window (general literature): Functional spectral studies commonly use pH ~6.5–8.0; extreme pH can denature or alter redox state.
• Ionic strength: Moderate ionic strength (e.g., 50–150 mM salt) stabilizes quaternary structure; very low ionic strength may promote aggregation.
• Redox control: Inclusion of mild reductants (e.g., dithionite for deoxy Hb generation) or controlled O2/CO/NO atmospheres is used in spectroscopic calibration; handle with appropriate safety and gas control.
• Additives: Avoid strong detergents/chaotropes unless intentionally denaturing. Glycerol (5–20%) can improve stability during cold storage; verify compatibility with your assay.

When to choose alternatives:

• For heme-peroxidase mimetics stable in organic co-solvents, consider purified hemin or horseradish peroxidase if assay tolerance to organics is required.
• For long-term room-temperature workflows, a lyophilized hemoglobin preparation may be preferable; reconstitute into your buffer of choice.

Note: Exact buffer composition and any preservative in this item are Not specified; refer to the CoA for compatibility considerations.

Storage & Reconstitution

Item-specific guidance from Product Data:

• Storage Conditions: Store at 2–8°C.
• Shipped In: Wet ice.

Additional practical guidance (general for aqueous hemoglobin solutions):

• Do not freeze unless the formulation is known to be freeze–thaw tolerant; freezing can cause aggregation and loss of activity/spectral integrity. If freezing is necessary, snap-freeze small aliquots in 10–20% glycerol and avoid repeated cycles.
• Protect from light to minimize photo-oxidation; store in amber or foil-wrapped containers.
• Minimize headspace and limit air exchange to reduce autoxidation; for oxyHb storage, air-equilibrated conditions are acceptable at 2–8°C for short periods. For deoxy or CO/NO forms, prepare fresh immediately before use.
• If particulates are observed, clarify by brief, cold centrifugation (e.g., 10,000 × g, 5 min) rather than filtration, which can adsorb protein.

Reconstitution: Not applicable — supplied as solution. If further dilution is required, use chilled, oxygen-appropriate buffer (composition matched to your assay). Buffer composition and any preservatives in this item are Not specified; refer to the CoA/Spec Sheet.

Stability: Without item-specific shelf-life data, use promptly upon receipt and monitor UV–Vis spectra over time to assess integrity. Always label aliquots with preparation date and state (oxy/deoxy/met).

Structure & Identity

Protein solution of hemoglobin at 10 mg/mL (research grade). Hemoglobin is a tetrameric heme protein; item-specific molecular identifiers are not provided for this listing.

• SKU: H1528851
• Product name: Hemoglobin Solution (10 mg/mL)
• Grade: BioReagent
• Concentration: 10 mg/mL (aqueous; exact buffer not specified)
• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey / SMILES: Not applicable to macromolecular protein mixtures; not specified.

Structural features (general, literature):

• Hemoglobin (e.g., adult HbA) is a heterotetramer composed of two α and two β globin chains, each noncovalently bound to a protoporphyrin IX heme cofactor carrying Fe(II).
• Quaternary structure exhibits cooperative O2 binding (T ↔ R allosteric transition) modulated by ligands (O2, CO, NO) and effectors (H+, CO2, 2,3-BPG; species-dependent).
• Each heme contains an iron atom coordinated to the porphyrin macrocycle and the proximal histidine of the globin; O2 binds at the sixth coordination site.

2D description in words (general):

• The protein scaffold forms a globular tetramer with internal heme pockets. The heme is a planar porphyrin ring with alternating double bonds (conjugated macrocycle), chelating Fe, with propionate side chains projecting toward solvent and hydrophobic edges buried within the globin fold.

Synthetic Utility

Not a small-molecule building block. Hemoglobin solution has limited direct role in synthetic organic chemistry. It is instead used as a biochemical model and catalyst mimic in aqueous systems.

Relevant to chemical research (general):

• Heme-centered catalysis studies: As a model for porphyrin–iron active sites, hemoglobin can inform the design of synthetic metalloporphyrin catalysts for oxygenation or peroxidation reactions.
• Conjugation chemistry: Surface lysines and cysteines (variant dependent) allow exploration of bioconjugation strategies (e.g., NHS-ester labeling, maleimide–thiol coupling) to study structure–function relationships.
• Ligand binding thermodynamics/kinetics: Provides a testbed for methods development (stopped-flow, rapid-freeze quench EPR/Mössbauer) applicable to synthetic heme analogs.

If your objective is preparative synthesis in organic media, consider switching to small-molecule heme models (e.g., hemin or iron porphyrins) that tolerate organic solvents and broader thermal ranges.

Target Specificity

This section typically applies to antibodies or affinity reagents and is not applicable to a hemoglobin protein solution.

• Antigen/epitope, clone, isotype, and species reactivity: Not applicable.

For experimental specificity, verify the hemoglobin variant/species from the CoA/Spec Sheet if your study requires a defined isoform.