Animal Cell Nuclear Separation Kit - BioReagent, Suitable for mammalian cell and tissue extract, 10-20T

Cat. No.: A777613

QUALITÀ & PUREZZA BioReagent, Suitable for mammalian cell and tissue extract, 10-20T

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Specifiche

Specifiche e purezza

BioReagent, Suitable for mammalian cell and tissue extract, 10-20T

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Grado

BioReagent, Suitable for mammalian cell and tissue extract

Product Line

Prodotti biologici

Application Protocols

Item-specific step-by-step instructions are not provided in the Product Data. The following is a generalized literature-style protocol for isolating nuclei from mammalian cells using differential lysis; adjust to your sample type and consult the kit insert/CoA for authoritative steps.

General workflow (overview):

1. Sample preparation: Harvest 1–10×10^6 cells (or equivalent tissue mass, minced on ice). Wash in cold isotonic buffer to remove media/serum.
2. Prechill: Place all buffers, tubes, and tools on ice. Add protease/phosphatase inhibitors freshly if required.
3. Cell lysis: Resuspend pellet in cold lysis buffer. Gently triturate or Dounce homogenize until most cells are lysed (monitor by trypan blue or phase contrast). Avoid over-homogenization.
4. Clarify: Low-speed spin to remove unbroken cells/debris. Collect supernatant containing released nuclei and cytosol.
5. Enrich nuclei: Pellet nuclei by appropriate centrifugation (RCF/time per insert). Carefully remove supernatant (cytosolic fraction). Wash pellet in cold wash buffer to improve purity.
6. Quality control: Assess nuclear integrity with DAPI/Hoechst staining; check purity by western blot for Lamin A/C (nuclear) vs GAPDH (cytosolic).
7. Downstream use: Proceed to ChIP, EMSA, ATAC-like nuclei prep, nuclear extract preparation, proteomics, or store nuclei briefly on ice prior to further processing.

Notes:

• Keep total processing time short and temperature cold.
• For tissue, optimize mechanical disruption and include nuclease/RNase considerations based on downstream assays.

All specific buffer volumes, centrifugation speeds, and incubation times are Not specified for this item; refer to the official kit insert.

Biological Roles

The kit itself has no intrinsic biological activity; it enables experimental access to the nuclear compartment of mammalian cells and tissues. Understanding nuclear biology can guide protocol choices and quality control.

Context (general biology):

• The nucleus houses genomic DNA organized into chromatin and is the site of DNA replication, most DNA repair pathways, and transcription of mRNA, rRNA (nucleolus), and many noncoding RNAs.
• Nuclear envelope integrity, nuclear lamina (lamins A/C, B), and nuclear pore complexes regulate nucleo-cytoplasmic transport and impact gene expression.
• Chromatin states (euchromatin vs heterochromatin) and histone post-translational modifications are sensitive to ionic strength, detergents, and temperature—factors controlled during nuclei isolation.

Experimental considerations:

• For studies of transcription factor binding or histone modifications, mild lysis and prompt processing at 0–4°C help preserve labile interactions (literature best practice).
• RNase-free conditions (DEPC-treated water alternatives, RNase inhibitors) may be required if nuclear RNA is a target; verify compatibility of inhibitors with the kit protocol.
• Validation markers: Lamin A/C and Histone H3/H4 (nuclear), GAPDH or Tubulin (cytosolic) for fractionation fidelity assessment by western blot or MS.

Note: The above are general principles. This item’s specific buffer composition and recommended handling are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

This kit’s buffers are intended for subcellular fractionation rather than classical solution buffering of biochemical reactions. Detailed compositions, pH values, and osmolarities are Not specified for this item; refer to CoA/Spec Sheet.

General/literature guidance for nuclear separation buffers (not item-specific):

• pH range: Neutral to slightly basic (approx. pH 7.2–8.0, literature) to maintain nuclear morphology and chromatin integrity.
• Ionic strength/osmolytes: Moderate salt (e.g., 100–250 mM ionic strength, literature) and osmolytes (e.g., sucrose) may be employed to stabilize nuclei during mechanical disruption and centrifugation.
• Detergents: Low concentrations of non-ionic detergents can permeabilize plasma membranes while preserving nuclear envelopes; minimize exposure to prevent nuclear rupture.
• Additives: Protease and phosphatase inhibitors are often used fresh. For RNA-focused applications, include RNase inhibitors and ensure RNase-free technique.

Practical tips:

• Prechill all buffers and tools; process samples on ice.
• Titrate lysis conditions by cell type: suspension vs adherent cells, soft vs fibrous tissues.
• Validate pH/osmolality after any user-added components to avoid osmotic shock.

If you need a buffer-only solution for general biochemical assays, consider purpose-built buffers with defined compositions rather than a nuclear separation kit.

Green Alternatives

Although this product is not a solvent or synthetic reagent, greener lab considerations apply to its use and disposal.

Sustainability perspective (general):

• Aqueous systems: Nuclear separation workflows are typically water-based, avoiding large volumes of hazardous organic solvents. This reduces VOC emissions and flammability risks compared with solvent-heavy extraction methods.
• Reduced single-use plastics: Where possible, opt for reusable, properly cleaned Dounce homogenizers and minimize disposable plastics (tubes, tips) without compromising sample integrity.
• Energy use: Consolidate prep sessions to reduce repeated freezer door openings and dry-ice consumption; store aliquots efficiently to limit freeze–thaw.
• Waste management: Segregate biological waste from general chemical waste. Avoid chlorinated disinfectants unless required; 70% ethanol or approved alternatives often suffice for area decontamination (institutional guidance prevails).

Comparison (general, literature-based):

• Differential lysis kits (aqueous) vs density gradients (sucrose/iodixanol):
  • Aqueous lysis buffers: Lower solvent hazard, faster, less waste; may require detergent cleanup for MS.
  • Density gradients: Often nonhazardous but higher material use and waste volume; can yield very pure nuclei with minimal detergents.

Procurement/packaging tips:

• Batch ordering to reduce shipment frequency and cold-chain materials.
• Request minimal/eco-friendly secondary packaging where possible.

Note: No item-specific green certifications are provided; consult the vendor documentation if needed.

Pharmaceutical Uses

This product is supplied strictly for research use only and is not intended for human or animal therapeutic, diagnostic, or clinical applications.

Contextual, non-clinical uses in pharmaceutical R&D (general):

• Mechanistic studies: Preparation of nuclear fractions from cultured mammalian cells or preclinical tissue models for target engagement assays, transcription factor occupancy profiling, or epigenetic mechanism studies.
• QC in discovery workflows: Assess nuclear vs cytoplasmic distribution of candidate compounds or CRISPR perturbations via western blot or proteomics following fractionation.
• Biomarker discovery: Nuclear proteome or chromatin modification mapping in preclinical research. Ensure downstream assay compatibility by removing salts/detergents as needed.

Regulatory note:

• No pharmacopeial monograph or GMP suitability is indicated for this kit. Use in regulated environments requires internal qualification, documented procedures, and verification of lot-specific CoA/SDS.

Any claims of clinical utility, diagnostic performance, or patient-related benefit are out of scope for this product and are not supported.

Physical Properties

Item-specific physicochemical specifications (for example, pH of each buffer, ionic strength, detergent content, and stabilizers) are not listed in the provided Product Data.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight/Formula: Not applicable to a multicomponent kit; Not specified for this item; refer to CoA/Spec Sheet.
• Boiling point, melting point, density, refractive index: Not applicable/Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Components are typically supplied as ready-to-use aqueous buffers and/or concentrates; exact compositions are Not specified for this item; refer to CoA/Spec Sheet.

Literature/general information (context, not item-specific):

• Nuclear extraction and separation buffers are commonly near neutral pH (≈7.2–8.0, literature) and formulated with salts (e.g., 100–250 mM ionic strength, literature) to maintain nuclear morphology during mechanical shearing and centrifugation.
• Mild non-ionic detergents are sometimes used at low percent levels to permeabilize plasma membranes while sparing nuclear envelopes (literature). Their exact identity and concentration in this item are not specified.

Practical notes:

• Keep buffers cold (on ice) during use to preserve nuclear integrity and minimize proteolysis (general best practice).
• If any components arrive as concentrates or lyophilized solids, reconstitute strictly per the kit insert and label aliquots to avoid freeze–thaw cycles.

For authoritative, item-specific properties (pH, composition, and stability), consult the CoA/Spec Sheet and SDS.

Quality and Grades

• Item-specific grade: BioReagent, Suitable for mammalian cell and tissue extract.

What this generally signifies (context for professional users):

• The reagents are formulated and quality-checked for research workflows involving mammalian cells/tissues, emphasizing preservation of nuclear structure and functionally relevant macromolecular complexes (chromatin-associated proteins, transcription machinery) during isolation. Exact QC tests for this item are not specified; consult CoA.
• BioReagent grade typically entails low bioburden, controlled pH/osmolality within target ranges, and performance testing on representative cell lines or tissues (literature/general). Specific acceptance criteria for this SKU are Not specified for this item; refer to CoA/Spec Sheet.

Implications for experimental success:

• Lot-to-lot consistency in buffer performance is important for reproducibility in downstream assays such as ChIP, EMSA, nuclear proteomics, and transcription factor binding studies. Verify lot data on the CoA.
• For sensitive readouts (mass spectrometry, enzyme kinetics), confirm whether any detergents, salts, or inhibitors present in the kit are compatible; remove or exchange buffers as needed (e.g., by gentle wash, spin filters, or dialysis).

Documentation to request/retain:

• CoA/Spec Sheet for each lot (item-specific performance notes, storage/stability windows).
• SDS for each component.
• Any stated compatibility notes for downstream applications (e.g., nuclease assays, MS).

Reaction and Applications

This is a workflow-oriented kit for biological separations rather than a chemical reagent for synthetic reactions. Typical research applications include nuclear enrichment for molecular biology, epigenetics, and proteomics.

Representative applications (general; expand according to your use case):

• Preparation of intact nuclei for chromatin immunoprecipitation (ChIP), CUT&RUN/CUT&Tag-style assays (after nuclei prep), and DNA–protein interaction studies such as EMSA.
• Isolation of nuclear fraction for transcription factor profiling, nuclear enzyme assays, immunoblotting (histones, lamin), and nuclear proteomics (following detergent/salt cleanup as needed).
• Subcellular fractionation to assess nuclear vs cytoplasmic localization by western blot or imaging (paired with cytosolic markers and nuclear markers such as Lamin A/C, Histone H3).
• Nuclei preparation from mammalian tissues for library preps that operate on isolated nuclei (e.g., single-nucleus omics approaches; validate compatibility with your platform).

Practical considerations (general best practices):

• Maintain cold conditions throughout to limit proteolysis and preserve protein–DNA interactions.
• Optimize lysis intensity for your cell type: gentle trituration or Dounce homogenization for soft tissues; brief detergent exposure for cultured cells. Over-lysis can rupture nuclei and release chromatin.
• Validate enrichment by microscopy (DAPI), flow cytometry of nuclei (DRAQ5/PI), and/or western blotting for nuclear and cytosolic markers.
• For downstream MS or nuclease-based assays, perform buffer exchange to remove interfering salts/detergents where necessary.

Note: This section summarizes common literature practices; refer to the item’s insert/CoA for kit-specific recommendations.

Reaction Conditions

The kit supports a biological workflow rather than a chemical reaction. Nevertheless, operational conditions critically affect outcome.

General operational conditions (literature/best practice; not item-specific):

• Temperature: 0–4°C throughout lysis and centrifugation to maintain nuclear integrity and minimize proteolysis.
• Time: Rapid processing (typically 15–45 minutes from lysis to first pellet, depending on sample and protocol) helps preserve labile complexes.
• Mechanical force: Gentle Dounce homogenization or pipette trituration for cells; adjust strokes based on cell/tissue toughness. Over-disruption can rupture nuclei.
• Centrifugation: Low to moderate g-forces for clearing debris and pelleting nuclei; specific speeds/RCF and durations must be taken from the kit insert (item-specific data are not provided here).
• Additives: Fresh protease/phosphatase inhibitors immediately before use; keep inhibitor solvents (e.g., DMSO) minimal to avoid membrane perturbation.

Quality control checkpoints:

• Microscopy: DAPI or Hoechst staining to confirm nuclear integrity and assess contamination.
• Marker analysis: Lamin A/C or Histone H3 for nuclear fraction; GAPDH, β-tubulin for cytosolic contamination.

Note: Exact volumes, times, and centrifugation parameters are Not specified for this item; refer to the kit’s CoA/Spec Sheet and insert for authoritative conditions.

Safety and Handling

Hazard classification details were not provided for this kit.

• Signal Word: Not specified for this item; refer to SDS.
• H-Statements: Not specified for this item; refer to SDS.
• GHS Classification/Pictograms: Not specified for this item; refer to SDS.

General laboratory safety guidance for biological reagent buffers (not item-specific; follow your institutional EHS policies):

• Personal protective equipment: lab coat, safety glasses, and appropriate chemical-resistant gloves. Use in a clean, designated area; avoid skin/eye contact and ingestion/inhalation of aerosols.
• Engineering controls: work on ice when required; use a biosafety cabinet if handling infectious or potentially biohazardous cell/tissue material.
• Incompatibilities: Avoid mixing with strong oxidizers or acids/bases unless specified by the protocol; do not autoclave unless the component label explicitly allows it (many buffers/detergents are heat-sensitive).
• First aid (overview; defer to SDS): rinse skin/eyes with water for several minutes upon contact; remove contaminated clothing. If inhaled as aerosol, move to fresh air. If ingested, rinse mouth; seek medical attention as per SDS.
• Spill response: Absorb aqueous spills with inert material; clean with water/detergent. Dispose of biological waste per local regulations.
• Temperature sensitivity: Store at −20°C as specified. Minimize repeated freeze–thaw cycles by aliquoting upon first thaw (good practice to preserve performance).

Always consult the kit’s SDS and institutional guidelines for definitive hazard and handling instructions.

Solvent Selection

This product is not an organic solvent; it is a kit of aqueous buffers and auxiliaries for subcellular fractionation. Traditional solvent selection parameters (polarity indexes, dielectric constants) are therefore not directly applicable.

Practical guidance (general, not item-specific):

• Matrix compatibility: The kit’s aqueous buffers are designed for mammalian cells and tissues. Avoid adding organic solvents (e.g., >5–10% v/v DMSO, ethanol) unless validated, as they may perturb membranes or chromatin compaction.
• Downstream assay compatibility: If performing mass spectrometry or enzyme assays sensitive to detergents/salts, consider a post-isolation buffer exchange into MS-compatible, low-salt buffers (e.g., 10–50 mM ammonium bicarbonate, literature) or detergent removal using spin columns.
• Additives: If your workflow requires protease/phosphatase/RNase inhibitors, confirm whether they are included; if not, add freshly prior to use, ensuring solvent vehicles (often DMSO) are kept minimal to avoid membrane effects (general best practice).

When to use this kit vs alternatives:

• Choose this kit when you need intact, enriched nuclei rapidly from mammalian samples with minimal cytoplasmic contamination for applications like ChIP, ATAC-like nuclei prep, EMSA, or nuclear proteomics.
• Consider density-gradient methods (e.g., sucrose/iodixanol, literature) when exceptionally high purity or special cell types (e.g., fragile primary neurons) demand gentler or more selective separations.

Storage and Reconstitution

• Storage Conditions (item-specific): Store at −20°C.
• Shipping (item-specific): Shipped in ice chest + ice pads to maintain the cold chain.
• Research Use Note (item-specific): For research use only.

General handling guidance for kit components (not item-specific; follow the kit insert):

• Upon receipt: Inspect components and immediately place at −20°C. If any components are labeled for 2–8°C, segregate accordingly (check the component labels/insert).
• Aliquoting: To minimize activity loss of labile ingredients, aliquot components upon first thaw into single-use portions. Avoid repeated freeze–thaw cycles.
• Reconstitution: If any components are supplied as concentrates or lyophilized powders, reconstitute with the specified volume and grade of water/buffer per the kit insert. Composition and volumes are Not specified for this item; refer to CoA/Spec Sheet.
• Stability: Working solutions kept on ice should be used the same day. Long-term stability claims are Not specified for this item; refer to CoA/Spec Sheet.
• Compatibility: Before adding user-supplied inhibitors (protease, phosphatase, RNase), confirm solvent vehicles and concentrations are compatible with nuclear integrity and downstream assays.
• Labeling and documentation: Mark aliquots with component name, lot, preparation date, and storage condition to ensure traceability.

Always defer to the kit’s official insert, CoA, and SDS for authoritative storage, reconstitution, and disposal instructions.

Structure and Identity

This product is a multicomponent life-science kit designed for subcellular fractionation of mammalian samples to isolate the nuclear fraction; it is not a single small molecule or polymer.

• SKU: A777613
• Product Name: Animal Cell Nuclear Separation Kit
• Category: Life Science reagent kit (研究/实验用)
• Grade/Purity: BioReagent, Suitable for mammalian cell and tissue extract (item-specific)
• Intended use: For research use only (item-specific)

Chemical identifiers (single-compound fields) are not applicable to a kit and are therefore not provided:

• Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not applicable for a multicomponent kit.
• InChIKey: Not applicable for a multicomponent kit.

Structural description (general):

• The kit typically comprises aqueous buffer systems and auxiliaries formulated to selectively disrupt plasma membranes while preserving nuclear integrity, followed by wash/cleanup steps to enrich intact nuclei. No specific component list is provided for this item.
• Functional design elements (general knowledge): hypotonic/iso-osmotic buffers, pH control near neutrality, salts for ionic strength, optional mild detergents for cytoplasmic release, and buffer additives that help maintain chromatin/nuclear envelope integrity during fractionation.

Note: Where specific identities and concentrations of kit components are required for regulated or validated workflows, consult the item’s Certificate of Analysis (CoA) and Specification Sheet.

Synthetic Utility

Not applicable. This product is a biological separation kit and is not used as a reagent or building block in chemical synthesis.

For users seeking chemical synthesis support:

• If your downstream work requires nuclei-derived biomolecules (e.g., DNA, histones) for conjugation chemistry, perform the biological isolation with this kit and then transition to chemical methodologies using appropriate, well-characterized reagents.
• For crosslinking or labeling within nuclei (e.g., NHS-ester dyes, photoaffinity probes), verify that any residual buffer components from the nuclear isolation are compatible with your chosen chemistry (for example, avoid primary amine-containing buffers when using NHS esters; general advice).

No named organic reactions, catalysts, or solvent systems are associated with the operation of this kit.

Target Specificity

This section typically applies to antibodies or affinity reagents. It is not applicable to a general nuclear separation kit.

• No antigen, clone, isotype, or species-reactivity data are associated with this SKU.

For experimental validation of nuclear enrichment, users commonly probe for:

• Nuclear markers: Lamin A/C, Histone H3/H4 (general literature practice).
• Cytosolic markers: GAPDH, β-tubulin (to assess purity).

These markers are not included or specified for this item unless stated in the kit components; consult the CoA/Spec Sheet.