Protein Lysis Buffer - BioReagent, sterile, 2×

Cat. No.: P1408622

GRADE & PURITY BioReagent, Sterile, 2×

Synonyms

RIPA Lysis Buffer | Lysis Buffer | Tissue Total Protein Lysis Buffer | SDS Lysis Buffer

Storage

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

Shipped In

Ice chest + Ice pads

Application

Protein Extraction

Overview

Product Introduction
This product is a high-performance protein lysis reagent launched by Aladdin with an optimized formula. It features simple operation and high lysis efficiency. When in use, mix it with the sample at a 1:1 ratio (volume ratio), and the lysed protein sample can be used in experiments such as BCA assay, Western Blot, and proteomics detection.
Operation Procedure
1.Experimental Preparation: Before the experiment, take the product out of the -20°C frozen environment to thaw. After the lysis buffer is completely melted and mixed evenly, place it on ice.
2.Add the sample and lysis buffer at a 1:1 volume ratio, mix thoroughly, and boil at 95°C for 10 minutes.
3.Centrifuge at 3000×g for 5 minutes, collect the supernatant, and use it for experiments including BCA protein concentration determination, Western Blot, and proteomics detection.
Precautions and Disclaimer
1.This product does not contain protease inhibitors. Add protease inhibitors by yourself according to experimental needs. Avoid repeated freezing and thawing; aliquoting for storage is recommended.
2.To ensure sufficient lysis, the protein concentration after lysis should be less than 2 μg/μL.
3.Store this reagent in the dark.
4.For your safety and health, wear a lab coat and disposable gloves during operation.
5.This product is limited to scientific research use by professional personnel. It shall not be used for clinical diagnosis or treatment, nor for food or pharmaceuticals.

Specifications

Synonyms

RIPA Lysis Buffer | Lysis Buffer | Tissue Total Protein Lysis Buffer | SDS Lysis Buffer

Specifications & Purity

BioReagent, sterile, 2×

Stability And Storage

Store at -20℃ long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle. Store in the dark.

Storage

Protected from light,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, Sterile

Application Protocols

Tested applications, recommended dilutions, and validated workflows specific to this catalog item are Not specified; refer to the product datasheet/CoA for any lot-specific instructions.

Note: General protocols for lysis and downstream use are provided under the Buffer Applications tab for literature-based guidance. Follow your lab’s SOPs and validate conditions with pilot experiments when adopting a new buffer formulation.

Biological Roles

This product is a laboratory buffer formulated for cell disruption and protein solubilization; it does not possess an intrinsic biological role in vivo.

General context (literature):

• Protein lysis buffers aim to preserve native protein conformation and post-translational modifications (PTMs) where required, while disrupting membranes and noncovalent interactions as needed. Buffer components are chosen to mimic physiological pH/ionic strength or to intentionally denature proteins for analytical workflows.
• Detergents form micelles that sequester lipids and hydrophobic protein segments, increasing solubility of membrane proteins. Chelators can inhibit metalloproteases; reducing agents maintain cysteine thiols; phosphatase inhibitors preserve phosphorylation states (if included—composition of this item is not specified).
• For functional assays, the choice of buffer affects enzyme activity, complex stability, and binding interactions. Therefore, the lysis buffer is a critical determinant of the biological integrity of extracted proteins but is not itself a biological effector.

No organism specificity, metabolic activity, or signaling capability applies to the buffer itself.

Buffer Applications

Protein Lysis Buffer is used to disrupt cells/tissues and extract proteins for downstream analyses. Exact composition for this item is Not specified; the following are general, literature-based practices.

Typical use scenarios:

• Total protein extraction for SDS-PAGE/Western blotting, ELISA, enzyme assays, IP/co-IP, and proteomics.

General procedure (literature example):

• Sample preparation: Pre-chill buffer and keep on ice. Harvest cells, wash with cold PBS, remove supernatant thoroughly.
• Lysis: Add ~5–10× pellet volume of buffer, pipette up/down or vortex briefly. Incubate 10–30 min on ice with occasional mixing. For tissues, homogenize mechanically.
• DNA shearing: Pass lysate through a narrow-gauge needle several times and/or sonicate briefly to reduce viscosity.
• Clarification: Centrifuge at 10,000–20,000 × g for 10–30 min at 4°C. Collect supernatant.
• Protein quantification: Choose an assay compatible with the buffer (e.g., BCA tolerates many detergents/reducing agents better than Bradford; verify with your exact formulation).

Compatibility notes:

• Affinity resins: Chelators (e.g., EDTA) can strip metal from IMAC resins; detergents may disrupt some protein–ligand interactions. Dialyze or desalt if necessary.
• Enzyme assays: Nonionic/zwitterionic detergents at low concentrations are often compatible; ionic detergents and strong chaotropes typically are not.

pH/salt ranges (literature, not item-specific): pH ~7.0–8.5; NaCl ~50–300 mM. Optimize for your protein system.

Green Alternatives

While aqueous protein lysis buffers are generally benign, environmental impact can arise from detergent choice, preservatives, and single-use plastics. Consider the following greener practices (literature/general):

• Detergent selection:
  • Prefer biodegradable, less toxic detergents (e.g., alkyl glucosides such as n-octyl-β-D-glucoside) over nonylphenol ethoxylates (e.g., legacy NP-40/Triton X-100), which are being phased out in many regions due to environmental persistence.
  • Use the minimum effective detergent concentration; leverage critical micelle concentration data to avoid excess.
• Preservatives and additives:
  • If sterility can be maintained, avoid azide-containing formulations where possible to mitigate heavy metal azide waste issues. If azide is present (not specified here), collect waste separately.
  • Choose TCEP over DTT where compatible; TCEP is odorless and more air-stable, potentially reducing waste.
• Process improvements:
  • Aliquot to reduce freeze–thaw waste (aligns with Product Data).
  • Implement detergent-removal cartridges/methods to allow downstream aqueous disposal of cleaned protein fractions where permitted by local regulations.
  • Reduce single-use plastics by using reusable, validated, autoclavable vessels for bulk preparations and cold-room workflows.

Trade-offs:

• Gentler or greener detergents may yield lower extraction efficiency for certain membrane proteins; compensate by optimizing salt, temperature (while keeping proteins stable), and extraction time.

Note: The specific composition of this item is Not specified; adapt green choices to your application.

Pharmaceutical Uses

This product is intended for research use only (Product Data) and is not formulated, qualified, or warranted for clinical or diagnostic applications.

Context in pharmaceutical R&D (general/literature):

• Preclinical research: Protein lysis buffers are ubiquitous in discovery biology for target validation, biomarker studies, and mechanism-of-action research.
• Process development: During upstream and downstream development, lysis buffers may be used to assess expression/solubility or for analytical characterization of cell banks and research lots. For GMP manufacturing or QC release testing, dedicated, validated reagents and SOPs are required.
• Excipient/formulation status: Lysis buffers are not pharmaceutical excipients; they are extraction reagents used on process intermediates or analytical samples. No pharmacopeial monograph applies to this product.

Regulatory cautions:

• Not for human or animal therapeutic use. Do not use in diagnostic procedures. For regulated workflows, verify composition, impurity profile, bioburden/endotoxin, and change-control provisions via formal quality agreements.

Item-specific details such as endotoxin level, preservative content, and GMP compliance are Not specified for this item; refer to CoA/Spec Sheet if required for risk assessments.

Physical Properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• pH, ionic strength, osmolarity: Not specified for this item; refer to CoA/Spec Sheet.
• Density, refractive index, conductivity: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Aqueous solution (general for lysis buffers). Specific miscibility/compatibility not specified for this item; refer to CoA/Spec Sheet.
• Boiling point, melting/freezing point: Not specified for this item; refer to CoA/Spec Sheet.

General/literature context for protein lysis buffers (not item-specific):

• Typical pH windows are near-neutral to slightly basic (pH ~7.0–8.5, literature) to balance protein stability with lysis efficacy.
• Common salt concentrations range from ~50–300 mM NaCl (literature), modulating protein solubility and minimizing nonspecific interactions.
• Detergent content can vary widely (e.g., 0.1–2% w/v or v/v, literature), with nonionic detergents often used to preserve protein activity, and stronger ionic detergents used for comprehensive solubilization.
• Temperature sensitivity: Many buffers are stored frozen to preserve sterility and inhibitor potency; repeated freeze–thaw cycles can degrade labile components (aligns with Product Data caution).

Note: For experimental design, measure the exact pH and detergent compatibility of the provided lot if critical to downstream assays (e.g., Bradford/BCA compatibility).

Quality & Grades

• Item-specific grade: BioReagent, Sterile (Product Data).

What this means:

• BioReagent: Indicates suitability for general biochemical research applications (e.g., protein extraction, enzymology, immunoassays). While high purity and low bioburden are typical, exact impurity limits are Not specified for this item; refer to CoA/Spec Sheet.
• Sterile: Manufactured and/or filtered to remove viable microorganisms. This supports applications where contamination would compromise results (e.g., cell-derived lysates for proteomics). Maintain sterility after first opening via aseptic handling and aliquoting.

Lot-specific quality attributes to consult on CoA/Spec Sheet (not provided here):

• pH and tolerance range at a defined temperature
• Endotoxin/bioburden levels, sterility test method
• Conductivity/ionic strength
• Detergent identity/concentration; presence of preservatives
• Protease/phosphatase inhibitor inclusion (if applicable)

Notes on stabilizers and UV background:

• If the formulation includes detergents or reducing agents, background absorbance at 280 nm and interference with colorimetric protein assays can occur (literature). Validate compatibility with your quantitation method.
• This item’s specific stabilizers and absorbance profile are Not specified for this item; refer to CoA/Spec Sheet.

Reaction & Applications

Primary application: lysis of cells and tissues to solubilize proteins for downstream biochemical analyses. No synthetic chemical reactions are intended.

Common research workflows (literature/general):

• Western blotting and SDS-PAGE: Robust extraction of total protein; ionic detergents denature proteins for size-based separation. If the buffer is nonionic/zwitterionic, SDS is added downstream during sample preparation.
• Immunoprecipitation/co-IP: Use milder, nonionic/zwitterionic detergent systems to preserve protein complexes; adjust salt to minimize nonspecific binding.
• Enzyme assays: Choose formulations without strong denaturants/reducing agents unless compatible with the enzyme of interest. Validate activity post-lysis.
• Proteomics: Employ MS-compatible buffers or perform cleanup (e.g., SP3, FASP, S-Trap) to remove detergents/salts prior to digestion and LC-MS.
• Nuclease management: Viscosity from genomic DNA can impede pipetting; shear mechanically (passaging or sonication) and/or add endonucleases (e.g., benzonase, if compatible) to reduce viscosity.

Practical tips:

• Keep samples cold (on ice/4°C) throughout to limit proteolysis and dephosphorylation. Add protease/phosphatase inhibitors freshly when required (if not pre-formulated; composition not specified for this item).
• Optimize buffer-to-pellet ratio (literature: ~5–10× pellet volume) and incubation time (10–30 min on ice) with gentle mixing.
• Clarify lysates by cold centrifugation to remove debris before analysis.

Note: Exact formulation and inhibitor content for this item are Not specified; consult CoA/Spec Sheet for lot details.

Reaction Conditions

As a lysis/extraction step rather than a chemical reaction, conditions emphasize temperature control, mixing, and mechanical disruption. The following are literature-based general guidelines (not item-specific):

• Temperature: Perform on ice or at 4°C to limit proteolysis and preserve PTMs.
• Time: Typical lysis incubation 10–30 min with intermittent mixing; tougher tissues may require longer or mechanical homogenization.
• Mechanical assistance: Pipetting, dounce homogenization, bead beating, or brief sonication to enhance disruption and reduce viscosity.
• Additives: Freshly add protease and phosphatase inhibitors if not pre-included (composition of this item is not specified). Consider DNase/benzonase to reduce viscosity if compatible with downstream assays.
• Clarification: Centrifuge at 10,000–20,000 × g for 10–30 min at 4°C to pellet debris; collect supernatant.
• Expected yield: Highly sample-dependent; membrane-rich tissues generally require stronger detergents or extended extraction. Evaluate visually (clarity) and analytically (protein assay) to optimize.

Compatibility reminders:

• Downstream electrophoresis: Ensure SDS and sample buffer are added for denaturing gels; remove incompatible detergents for native gels.
• Immunocapture: Use milder detergents and moderate salt to preserve complexes; avoid chelators when using metal-dependent interactions.

Exact conditions and composition for this item are Not specified; consult CoA/Spec Sheet and tailor to your sample type.

Safety & Handling

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

General safety guidance for protein lysis buffers (literature/general knowledge):

• Potential hazards depend on formulation. Detergents/surfactants can cause skin/eye irritation; reducing agents or chaotropes (if present) may be corrosive or harmful. Handle with gloves, lab coat, and safety glasses. Work in a fume hood if aerosols or volatile additives are present.
• Avoid inhalation and contact with eyes/skin. In case of exposure, rinse affected area with copious water and seek medical attention as appropriate. If ingested, rinse mouth; do not induce vomiting; consult medical services.
• Prevent microbial contamination during use to maintain sterility (Product Data: Sterile). Use aseptic technique when opening and aliquoting.
• Storage incompatibilities: Do not mix unknown formulations with strong oxidizers or acids/bases without compatibility verification. If the buffer contains sodium azide (common in some sterile buffers; composition not specified here), avoid contact with acids and copper/lead plumbing due to azide’s reactivity (literature caution; verify SDS).
• Spill response: Absorb with inert material, collect for disposal, and rinse area. Detergent-containing spills can be slippery—mitigate slip hazards.

Always defer to the product’s SDS and your institution’s safety protocols for PPE, exposure controls, and disposal.

Solvent Selection

This product is an aqueous buffer; organic solvent selection is not typically applicable. However, choosing among aqueous lysis systems involves selecting detergent class and ionic conditions appropriate for your target proteins.

General guidance (literature):

• Polarity/matrix: Highly polar, water-based. Dielectric constant reflects water; additives modulate micelle formation and protein solubility.
• Detergent classes and typical use-cases:
  • Nonionic (e.g., Triton-like, octyl glucoside): Gentler, preserve activity and complexes; good for membrane proteins at modest concentrations.
  • Zwitterionic (e.g., CHAPS): Balance between extraction strength and native-state preservation; compatible with some MS workflows after cleanup.
  • Ionic (e.g., SDS): Strong solubilization/denaturation; suited for SDS-PAGE workflows but disrupts native interactions.
• Salt and buffer choice:
  • Moderate ionic strength (~50–300 mM NaCl, literature) reduces nonspecific binding; buffer pH near neutrality (pH ~7–8.5, literature) supports stability.
• Additive compatibility:
  • Chelators (e.g., EDTA) inhibit metalloproteases but can disrupt metal-dependent complexes and affinity resins.
  • Reducing agents (e.g., DTT, TCEP) maintain thiols but can interfere with maleimide chemistries; TCEP is more air-stable and MS-friendly than DTT.

For this specific item, exact detergent/ion composition is Not specified; refer to CoA/Spec Sheet. Select downstream-compatible cleanup (desalting, detergent removal) as needed.

Storage & Reconstitution

• Storage (Product Data):
  • Protect from light.
  • Store at −20°C.
  • Avoid repeated freezing and thawing.
  • Shipped in ice chest with ice pads to maintain cold chain.

Best practices (general):

• Upon receipt: Verify container integrity and immediately store at −20°C as directed.
• Aliquoting: To minimize degradation and maintain sterility (Product Data: Sterile), thaw once on ice, mix gently, and dispense into sterile, low-protein-binding tubes in single-use aliquots. Refreeze promptly at −20°C or colder as appropriate.
• Thawing: Thaw aliquots on ice. Mix by gentle inversion. If any reversible precipitation is observed after thaw (can occur with some detergents/salts), warm briefly to room temperature and mix until clear; do not overheat.
• Light protection: Keep in amber/foil-wrapped containers or a dark storage box to protect light-sensitive components (if any; formulation not specified).
• Reconstitution: Not applicable if supplied as a ready-to-use liquid. If provided frozen/partially solid, allow complete thaw and homogenization before use. Any concentrate/dilution instructions are Not specified for this item; refer to CoA/Spec Sheet.
• Freeze–thaw: Avoid cycling; repeated freeze–thaw can degrade labile inhibitors and alter detergent performance.

Dispose of unused material according to institutional and local regulations, especially if preservatives are present (composition not specified).

Structure & Identity

• This product is an aqueous mixture formulated for efficient disruption of cells and solubilization of proteins (protein lysis buffer). It is not a single defined small molecule.

• Item-specific identifiers:

  • SKU: P1408622
  • Product Name: Protein Lysis Buffer
  • Grade/Purity: BioReagent, Sterile (Product Data)

• Composition identifiers:

  • 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.
  • SMILES / InChIKey: Not applicable to complex mixtures; not specified for this item; refer to CoA/Spec Sheet.

• Structural features (general knowledge):

  • Typical protein lysis buffers are aqueous solutions containing buffering agents (e.g., Tris, HEPES), salts (e.g., NaCl), and one or more surfactants/detergents (e.g., nonionic or zwitterionic) to solubilize membranes and proteins. They may optionally include chelators (e.g., EDTA), reducing agents, or enzyme inhibitors; actual components for this item are not specified.

• 2D structure description:

  • Not applicable; the product is a multi-component solution rather than a discrete molecular structure.

Synthetic Utility

Not typically applicable. Protein Lysis Buffer is not a reagent for organic synthesis or small-molecule transformations.

General laboratory interface (literature):

• Bioconjugation workflows: Detergents, high salt, chelators, and reducing agents commonly present in lysis buffers can inhibit downstream chemical modifications (e.g., NHS-ester or maleimide labeling) and affinity purifications. If conjugation is planned, perform buffer exchange (spin columns, dialysis) into compatible media (e.g., PBS without amines for NHS-ester reactions; remove reductants before maleimide–thiol coupling).
• Enzymatic digestions: For proteomics, remove or dilute detergents and salts that suppress trypsin activity and MS ionization; adopt MS-friendly surfactants or cleanup methods (e.g., S-Trap, SP3) as needed.

No named synthetic reactions, catalysts, or stoichiometric transformations pertain to this product.

Target Specificity

Not applicable. Protein Lysis Buffer is a general-purpose extraction reagent and does not have target specificity, antigen/epitope recognition, or clone/isotype characteristics.

Item-specific target reactivity data are Not specified for this item; refer to CoA/Spec Sheet if any application-specific validations are provided.