Hemoglobin Solution (2%) - BioReagent

Cat. No.: H1528875

GRADE & PURITY BioReagent

Storage

Store at 2-8°C

Shipped In

Wet ice

Overview

2% Hemoglobin Solution is an aqueous solution prepared from animal-derived hemoglobin, and is a commonly used reagent in biochemical, immunological, and microbiological experiments. It can be used in enzyme activity assays, substrate reaction systems, hemolysis-related experiments, complement fixation tests, microbial blood agar-related tests, and protein reference experiments. Note: This product is a biological solution with potential biological risks. Wear gloves during handling, avoid skin and mucous membrane contact, and avoid inhalation. For research use only. Not for clinical diagnosis or human administration.

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

No item-specific tested applications are provided in the Product Data. The following generalized protocols are offered as starting points (literature-based; adjust to your lab’s requirements). Always verify compatibility with the item’s buffer and concentration.

• A. UV–Vis characterization of hemoglobin state (general)

  • Materials: Hemoglobin Solution (2%), compatible buffer (pH ~7.4), quartz cuvettes.
  • Procedure: Equilibrate samples at 20–25 °C. Dilute an aliquot to yield an Amax between 0.2–1.0 at the Soret band (typically 405–420 nm depending on redox/ligand state). Record spectra 250–700 nm. Assign state by band positions/intensities per literature. Note: Extinction coefficients and exact band positions are species/state dependent.

• B. Pseudoperoxidase activity assay with TMB (general)

  • Prepare acetate or citrate-phosphate buffer (pH 4.5–5.0). Dilute hemoglobin to low µM range. In a cuvette or microplate, add TMB working solution, initiate with H2O2 (e.g., 100–300 µM), and monitor increase at 652 nm. Include blank (no Hb) and H2O2-only controls. Stop reaction by acidification if desired. Optimize to balance signal and protein stability.

• C. Oxygen-binding demonstration (general)

  • Deoxygenate a diluted Hb solution by N2 sparging. Record deoxy spectrum. Re-equilibrate with air or O2-enriched gas and record oxy spectrum. Optionally, generate CO-Hb in a fume hood with trace CO for comparison. Handle gases safely and confirm states spectrally.

These are illustrative workflows; for regulatory or quantitative purposes, establish validated SOPs. Item-specific parameters are Not specified for this item; refer to CoA/Spec Sheet.

Biological Roles

General literature overview (not item-specific; source organism for this product is not specified):

• Core function

  • Hemoglobin (Hb) is a tetrameric heme protein responsible for reversible oxygen transport in many organisms. Each subunit binds one O2 molecule at the ferrous heme iron, enabling cooperative binding and efficient O2 delivery.

• Allostery and regulation

  • Exhibits cooperative binding (sigmoidal O2 saturation curve) and the Bohr effect, where decreased pH and increased CO2 lower O2 affinity. Organic phosphates (e.g., 2,3-BPG in many vertebrates) modulate affinity by stabilizing the deoxy (T) state.

• Redox and ligand chemistry

  • The heme iron cycles among Fe(II) (oxygen-binding competent), Fe(III) (methemoglobin, non-O2-binding), and high-valent intermediates during peroxidatic reactions. Competing ligands include CO and NO with distinct affinities, useful in mechanistic studies.

• Structural biology

  • The globin fold protects the heme and tunes ligand access via the distal pocket. Subunit interfaces transmit conformational changes during T ↔ R transitions, underpinning cooperative binding.

• Biotechnological utility

  • Serves as a model for protein–ligand interactions, allosteric regulation, and redox enzymology; used in educational labs to illustrate spectroscopy, gas binding, and protein denaturation/renaturation phenomena.

Note: The above describes hemoglobin generally. For this catalog item, specific biological source, post-translational modifications, and ligand/oxidation state are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

This product is a protein solution and is not a buffering reagent. Therefore, classical buffer system recipes (e.g., phosphate, Tris, HEPES) do not apply directly.

• Practical notes (general)
  • Use compatible biological buffers (typically pH ~7–8) to dilute or formulate working solutions to maintain hemoglobin structural integrity and function.
  • Avoid extreme pH or denaturing additives when preparing assay media.

For detailed buffer composition that accompanies this specific item (if any), Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

While this is a biological aqueous solution rather than an organic solvent or synthetic reagent, greener practice considerations still apply.

• Greener handling choices (general)

  • Prefer aqueous buffers at neutral pH and avoid halogenated solvents entirely; this reagent is inherently water-based.
  • Right-size aliquots to reduce waste from expired material and minimize cold-chain energy from repeated shipments.
  • Use recyclable, low-adsorption plastics where feasible and follow local regulations for nonhazardous aqueous biological waste.

• Alternative materials to consider (context-dependent)

  • For heme-catalyzed oxidation in organic media: synthetic metalloporphyrins in green solvents (e.g., ethanol or ethyl acetate where appropriate) can obviate biological material use; trade-off is reduced biological relevance.
  • For teaching demonstrations requiring color change but not biological proteins: iron porphyrin model compounds or plant-derived heme proteins (e.g., leghemoglobin) may provide renewable alternatives (availability varies).

• Comparison snapshot (general)

  • This item: Ready-to-use aqueous protein, no volatile organic compounds, cold-chain required (2–8 °C), potential biological origin.
  • Synthetic heme models: Dry, often room-temperature stable, solvent-selectable, but may necessitate organic solvents and lack cooperative binding phenomena.

Overall, the supplied aqueous format already meets many green chemistry principles (benign solvent, low VOC). Focus on minimizing cold storage/transport footprints and right-sizing usage to reduce waste.

Pharmaceutical Uses

This BioReagent-grade hemoglobin solution is intended for research use only (from Product Data) and is not supplied with documentation for pharmaceutical manufacturing or clinical diagnostic use.

• Non-therapeutic laboratory roles (general)

  • Reference material for assay development, method validation, and instrument calibration in research settings.
  • Model protein for formulation studies (e.g., assessing protein stability, aggregation) in preclinical research.

• Not included/specifically not claimed for this item

  • GMP compliance, pharmacopeial monograph compliance, lot release testing to clinical standards, or validated use as an in vitro diagnostic control. These attributes are Not specified for this item; refer to CoA/Spec Sheet if needed.

Any use in regulated contexts would require independent qualification/validation by the end user and appropriate sourcing; Aladdin’s listing specifies For research use only.

Physical Properties

Item-specific specifications not listed below are not provided by the Product Data.

• Provided for this item

  • Form: Aqueous protein solution
  • Nominal concentration: 2% (from product name; basis not specified)

• Not specified for this item; refer to CoA/Spec Sheet

  • Exact buffer composition, pH, ionic strength
  • Protein source (species), extinction coefficient, viscosity, osmolality
  • Density, refractive index, conductivity

• General/literature properties of hemoglobin (for context; not item-specific)

  • Approx. molecular mass (tetramer): ~64.5 kDa; monomer (globin) ~16 kDa.
  • Isoelectric point (pI): typically ~6.7–7.1 depending on species and modification.
  • UV–Vis spectral features: intense Soret band ~405–420 nm (met/deoxy/oxy forms differ), Q-bands in visible region; CO/dioxygen difference spectra used analytically.
  • Oxygen binding: cooperative (Hill coefficient >1; literature often ~2.6–3 at 25 °C in physiological buffers for adult Hb). Sensitive to pH, ionic strength, and allosteric effectors (e.g., phosphate/organophosphates; species-dependent).
  • Stability: Heme proteins are sensitive to oxidation (Fe2+ → Fe3+, methemoglobin) and denaturation at elevated temperatures or extreme pH.

• Solubility (general)

  • Freely soluble in physiological aqueous buffers; insoluble in nonpolar organic solvents. High salt or extreme pH can precipitate/denature globular proteins.

Use these literature values only for conceptual planning; rely on the item’s CoA/SDS for exact specifications.

Quality and Grades

• Item-specific grade

  • Grade: BioReagent (from Product Data)

• What BioReagent grade typically implies (general explanation)

  • Suitable for common biological research applications such as enzymology, protein chemistry, and routine biochemical assays.
  • Emphasis on functional integrity and low levels of interfering contaminants for typical bioassays, but not necessarily certified for clinical diagnostics or GMP manufacturing.

• Considerations specific to hemoglobin solutions

  • Oxidation state matters: Functional studies often require ferrous (oxy-/deoxy-) hemoglobin, whereas ferric (methemoglobin) has distinct spectra/activities. The provided Product Data do not specify oxidation state; verify via UV–Vis or supplier CoA prior to critical experiments.
  • Additives/preservatives: Presence of stabilizers (e.g., salts, buffers, antimicrobial agents) can affect downstream assays (peroxidase-like activity, binding studies). Not specified for this item; refer to CoA/Spec Sheet.

• Typical QC assays (illustrative; not item-specific)

  • UV–Vis spectrum with defined Soret/Q-band ratios; SDS-PAGE for purity/aggregation; total protein determination (e.g., A280 or colorimetric); methemoglobin percentage by spectral deconvolution.

• Documentation

  • For regulated or highly quantitative applications, request the lot-specific Certificate of Analysis (CoA) detailing concentration basis (w/v vs. protein content), buffer composition, and acceptance criteria. Where parameters are not provided in the Product Data, treat them as Not specified for this item; refer to CoA/Spec Sheet.

Reaction and Applications

As a biological macromolecule, hemoglobin is not a conventional reagent for organic synthesis. Its utility lies in biochemical and biophysical applications. The Product Data list no specific applications; below are common research uses (general literature guidance):

• Spectrophotometric standards and teaching demonstrations

  • Use the Soret and visible bands to teach ligand binding (O2/CO/NO) and redox interconversions (oxy ↔ deoxy ↔ methemoglobin) by UV–Vis.

• Model peroxidase-like catalysis (pseudoperoxidase activity)

  • Hemoglobin can catalyze H2O2-dependent oxidation of chromogenic substrates (e.g., TMB, ABTS) via the heme center (general literature). Conditions must be optimized to avoid protein damage.

• Oxygen-binding and allostery studies

  • Measure O2 dissociation curves and Hill coefficients under varying pH/ionic strength to illustrate Bohr effect and cooperative binding (general).

• Calibration/control material for colorimetric hemoglobin assays

  • In vitro assay development and instrument verification (e.g., cyanmethemoglobin or alternative chemistries) in research labs. Note: Not for clinical diagnostics without appropriate validation.

• Protein chemistry workflows

  • Reference protein for SDS-PAGE banding (~16 kDa monomer under denaturing conditions), UV–Vis method validation, and heme quantification protocols (literature).

• Practical tips (general)

  • Maintain at 2–8 °C; minimize light during redox-sensitive assays.
  • Pre-equilibrate with gas mixtures (N2/O2/CO) using gas-tight syringes or tonometers for binding studies; confirm state by spectra.
  • For peroxidase-like assays, use low micromolar protein and submillimolar H2O2 to balance signal and stability (optimize empirically).

Reaction Conditions

Classical synthetic “reaction conditions” are not applicable. For laboratory studies involving hemoglobin’s biochemical activities, the following general literature guidance may help design experiments (not item-specific; optimize empirically):

• Spectrophotometric characterization

  • Buffer: PBS, HEPES, or Tris, pH 7.0–7.5. Temperature: 20–25 °C. Record UV–Vis spectra 250–700 nm. Minimize light exposure during redox studies.

• Oxygen/ligand binding assays

  • Use tonometry or gas equilibration in sealed cuvettes. Prepare deoxy-Hb by gentle deoxygenation under N2/argon; for CO-binding spectra, equilibrate with low % CO in carrier gas in a fume hood. Confirm state by characteristic spectra (Soret/Q-band positions; literature).

• Pseudoperoxidase activity assays

  • Substrate: e.g., TMB or ABTS (chromogenic), H2O2 as oxidant. Buffer: citrate-phosphate or acetate (pH 4–6) for higher peroxidatic rates, or neutral pH for milder conditions. Typical ranges: Hb low µM; H2O2 50–500 µM. Monitor at substrate-specific wavelengths (e.g., 652 nm for TMB diimine). Quench promptly to limit protein damage.

• Protein handling

  • Keep on ice during setup; pre-chill buffers. Use low-binding plastics. Filter or centrifuge gently if aggregates are observed.

Exact concentrations, extinction coefficients, and buffer composition for this product are Not specified for this item; refer to CoA/Spec Sheet and SDS.

Safety and Handling

• GHS/SDS information for this item

  • Signal word, hazard statements, classification, pictograms: Not specified for this item; refer to SDS.

• General safety guidance for protein solutions (non-hazard-specific)

  • PPE: Wear lab coat, safety glasses, and appropriate disposable gloves. Avoid aerosol generation and splashes.
  • Biosafety considerations: If of biological origin, handle as a potential bio-derived material of unknown infectious status unless supplier documentation states otherwise. Work on a clean bench; disinfect surfaces after use.
  • Incompatibilities: Avoid strong oxidizers (can convert heme Fe2+ to Fe3+), strong acids/bases (denaturation), and metal chelators that may perturb heme iron coordination. Avoid contact with hypochlorite in presence of protein to limit chlorination byproducts.
  • Environmental precautions: Prevent release to drains if local regulations restrict disposal of proteinaceous/biological materials.

• First-aid (general, defer to SDS)

  • Inhalation: Move to fresh air if aerosols were generated; seek medical advice if symptoms persist.
  • Skin/eye contact: Rinse with water for several minutes. Remove contaminated clothing/gloves. Obtain medical attention if irritation occurs.
  • Ingestion: Rinse mouth. Do not induce vomiting. Seek medical advice.

• Handling and use tips

  • Keep cold (2–8 °C as specified) and minimize time at ambient temperature.
  • Avoid repeated freeze–thaw; if freezing is contemplated, confirm compatibility with supplier guidance (not specified for this item).
  • Work with clean, low-protein-binding plastics or silanized glassware to minimize adsorption.

Solvent Selection

This product is supplied as an aqueous 2% protein solution and is used in aqueous systems; traditional organic solvent selection is generally not applicable.

• Applicable diluents (general guidance)

  • Physiological buffers: Phosphate-buffered saline (PBS, pH ~7.2–7.6), HEPES, or Tris buffers (pH ~7–8) are commonly compatible with globular proteins, depending on the intended assay.
  • Deoxygenated buffers (for deoxy-Hb work): Degas by nitrogen/argon sparging; include reducing agents only if compatible with your assay.
  • Avoid: Strongly acidic/basic buffers, high concentrations of chaotropes, or organic cosolvents that denature proteins (e.g., >20–30% v/v ethanol, acetone, acetonitrile).

• Polarity/miscibility (general)

  • Hemoglobin is water-soluble; the solution is fully miscible with water and typical aqueous buffers. It is not soluble in nonpolar organic media.

• When to choose this reagent vs alternatives

  • Use an aqueous hemoglobin solution when a ready-to-use heme protein standard or model is desired for spectrophotometry, peroxidase-like activity studies, or oxygen-binding demonstrations without needing protein solubilization steps.
  • If organic-tolerant heme catalysts are needed, consider hemoprotein mimics or synthetic metalloporphyrins (general alternative, not directly comparable to this biological reagent).

• Compatibility notes

  • Metal chelators (EDTA) and high phosphate can alter ligand binding equilibria; evaluate effects empirically for your assay.
  • Ionic strength and pH near the protein’s pI may promote aggregation; maintain moderate ionic strength (~100–200 mM, literature guidance) to enhance colloidal stability.

Storage and Reconstitution

• Storage conditions (from Product Data)

  • Store at 2–8 °C.
  • Shipped on wet ice.

• Reconstitution

  • Not applicable: the product is supplied as a ready-to-use solution. If concentration adjustment is required, dilute with a compatible buffer appropriate for your assay (buffer composition and pH are Not specified for this item; refer to CoA/Spec Sheet).

• Handling and aliquoting (general best practices)

  • Upon receipt, inspect for container integrity and any signs of precipitation. Mix gently by inversion; avoid vigorous vortexing or foaming.
  • Prepare single-use aliquots in low-protein-binding tubes to minimize freeze–thaw and adsorption losses.
  • Protect from prolonged light if redox state is critical to your work.

• Stability notes

  • Avoid repeated freeze–thaw cycles. If longer-term storage is needed and freezing is contemplated, first confirm compatibility with the product’s buffer/preservatives (not specified) and consider adding cryoprotectants only if suitable for your application.
  • Monitor function/identity over time by UV–Vis spectral checks (Soret/Q-band) to assess oxidation or denaturation.

• Research use note

  • For research use only (from Product Data). Defer to the product’s SDS and lot-specific CoA for definitive stability and compatibility information.

Structure and Identity

Hemoglobin Solution (2%) is a ready-to-use aqueous protein preparation of the oxygen-binding metalloprotein hemoglobin for biochemical research workflows.

• Item identity (from Product Data)

  • Product: Hemoglobin Solution (2%)
  • Grade: BioReagent
  • Physical form: Aqueous solution (protein), 2% stated by product name
  • CAS, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.

• Macromolecular characteristics (general literature, not item-specific)

  • Hemoglobin is a tetrameric globular protein composed of four polypeptide subunits, each noncovalently bound to a heme (protoporphyrin IX) containing Fe(II).
  • Approximate tetrameric molecular mass: ~64–65 kDa (literature; varies by species and modification state).
  • Functional groups/features: porphyrin macrocycle, iron(II) coordination sphere (proximal histidine ligand), multiple ionizable side chains; no single small-molecule formula is applicable.
  • Quaternary structure: two α-like and two β-like subunits (literature) with cooperative O2 binding.

• 2D structural description (general)

  • Protein backbone with α-helical content forming globin folds; each subunit encloses a planar heme prosthetic group with a central Fe(II). The iron coordinates to four porphyrin nitrogens, a proximal histidine from the protein, and binds O2 at the sixth coordination site under oxygenated conditions.

• Notes on identity confirmation (general guidance)

  • Typical identity checks include UV–Vis spectral features of heme proteins (Soret band), SDS-PAGE protein banding (~16 kDa per monomer; tetramer dissociates under denaturing conditions), and CO/O2 difference spectra. Specific acceptance criteria are item-dependent (not provided here).

Synthetic Utility

This reagent is a biological macromolecule and is not a building block or reagent for classical organic synthesis.

• Not typically applicable

  • It is not used for bond-forming reactions (e.g., cross-coupling, carbonyl chemistry) or as a protecting group reagent.

• Where it can be useful (general, biochemical context)

  • As a catalytic model: heme center exhibits peroxidase-like activity useful for studying redox mechanisms under aqueous conditions.
  • As a spectroscopic standard: defined heme absorbance bands enable instrument qualification and method development.

For conventional synthetic applications, consider small-molecule heme analogs or metalloporphyrins, which offer greater solvent compatibility and well-defined stoichiometry.

Target Specificity

Not an antibody or affinity reagent; target specificity does not apply.

• This product is a hemoglobin protein solution intended as a general biochemical reagent/standard. No antigen/epitope, species reactivity, clone, or isotype information is relevant.

Any affinity interactions (e.g., ligand binding) discussed elsewhere refer to general hemoglobin biochemistry, not to selective binding akin to immunoreagents.