pLenti-CBLL2-sgRNA

Cat. No.: P744455

Specifications

Product Name

pLenti-CBLL2-sgRNA

Product Line

Produits biologiques

Stockage et expédition

Conditions de stockage de stockage

Store at -80°C

Expédié en

Dry ice packs + Cold packs

Stabilité et stockage

Stored at -20 ℃, valid for 3 months;Stored at -80 ℃, valid for at least one year.

Application Protocols

No tested application protocols, recommended dilutions, or instrument settings are provided in the Product Data. For research use, employ institutionally approved protocols for handling plasmid DNA and, where applicable, for producing/using lentiviral vectors. Validate performance in your specific system with appropriate controls (non‑targeting and positive control sgRNAs). Refer to CoA/Spec Sheet for any item‑specific handling notes if available.

Biological Roles

• Item-derived target: CBLL2 (Cbl proto‑oncogene like 2). The product name indicates the sgRNA is designed to direct CRISPR machinery to CBLL2; spacer details and exact isoform targeting are not provided in the Product Data.
• Gene/function overview (literature/general, not item‑specific): CBLL2 encodes a protein with homology to Cbl family adaptor/E3 ubiquitin ligases implicated in protein ubiquitylation and endocytic trafficking. While CBLL1 (HAKAI) is well characterized as an E3 ligase for E‑cadherin, CBLL2 is considered a related gene with roles suggested in signal transduction pathways and post‑translational regulation. Functional redundancy/compensation among Cbl‑like proteins has been discussed in the literature, but precise roles of CBLL2 can be context dependent.
• Research implications (general): Targeted perturbation of CBLL2 enables studies of ubiquitin‑mediated regulation, receptor turnover, and signaling crosstalk. Phenotypic outcomes may vary by cell type, expression level, and presence of paralogs.
• Controls and verification (general): Employ multiple independent sgRNAs to deconvolute on‑target from off‑target effects. Validate gene perturbation at the DNA (indel profiling), RNA (qPCR, RNA‑seq), and protein (immunoblot, proteomics) levels. Note: No biological activity, clinical, or diagnostic claims are made for this product. It is for research use only.

Buffer Applications

Not typically applicable. Plasmid DNA is commonly supplied in simple aqueous buffers (e.g., nuclease‑free water or low‑salt TE) rather than used as a buffering agent. If the supplied buffer composition affects your downstream assay (e.g., Mg2+ sensitivity for certain enzymes), consult the CoA/Spec Sheet and adjust by dialysis or spin‑column exchange as appropriate to your institutional protocols (general guidance, not item‑specific).

Green Alternatives

Not directly applicable. This is a plasmid DNA reagent rather than a solvent or consumable with conventional green-chemistry alternatives. Sustainability considerations (general):

• Minimize single‑use plastics by planning aliquots and avoiding repeated freeze–thaw.
• Use enzymatic cleanup kits and aqueous workflows that reduce organic solvent use during plasmid preparation and downstream analysis (literature/general).
• Consolidate shipments and maintain the cold chain to avoid material loss and reshipments.

Pharmaceutical Uses

Not applicable. This reagent is for research use only and is not intended for pharmaceutical manufacturing, clinical, or diagnostic use. No pharmacopeial status or excipient role is provided in the Product Data. Do not use in human or veterinary applications.

Physical Properties

• Physical state/appearance — Not specified for this item; refer to CoA/Spec Sheet. Plasmid DNA is typically supplied in aqueous buffer or lyophilized (literature/general).
• Molecular weight — Not specified for this item; refer to CoA/Spec Sheet. (Literature/general: pLenti backbones commonly range ~7–12 kb, corresponding to ~4.6–7.4 MDa for the plasmid circle.)
• Solubility (literature/general): Highly soluble in aqueous, low‑salt buffers (e.g., TE, nuclease‑free water). Insoluble in most organic solvents.
• pKa/logP — Not applicable for nucleic acid polymers (no single pKa/logP; behavior dictated by the polyanionic phosphate backbone) (literature/general).
• UV/Vis (literature/general): DNA exhibits a strong absorbance peak at ~260 nm; A260 of 1.0 ≈ ~50 µg/mL for dsDNA (pathlength 1 cm). Hyperchromicity upon denaturation.
• Density, refractive index, mp/bp — Not applicable/reported for plasmid DNA in aqueous solution; Not specified for this item; refer to CoA/Spec Sheet.
• Stability (general): Sensitive to nucleases, repeated freeze–thaw, and elevated temperatures; divalent cations and high pH can accelerate degradation. Always handle with nuclease‑free consumables. Note: All numerical values above, where given, are literature/general and not item‑specific unless explicitly stated.

Quality and Grades

• Grade/purity — Not specified for this item; refer to CoA/Spec Sheet.
• Typical quality attributes for plasmid DNA (literature/general):
  • Sequence identity of the sgRNA spacer and vector backbone verified by Sanger sequencing.
  • High supercoiled fraction; low levels of genomic DNA, RNA, endotoxin, and residual salts/solvents depending on the intended downstream use.
  • A260/280 ~1.8–2.0 and A260/230 >2.0 (for highly purified DNA), when applicable.
• Endotoxin level (if relevant for cell work) — Not specified for this item; refer to CoA/Spec Sheet.
• Formulation (buffer, concentration) — Not specified for this item; refer to CoA/Spec Sheet.
• What this means practically: In the absence of item‑specific grade claims, users should confirm suitability for their workflow (e.g., cloning, transfection, or packaging) by reviewing the CoA/Spec Sheet and, if necessary, performing small‑scale pilot tests.
• Documentation: For regulated environments, request sequence maps, antibiotic resistance details, and restriction digest patterns as applicable. Note: No stabilizers, antibiotics, or excipients are claimed in the Product Data; do not assume their presence—verify on the CoA/Spec Sheet.

Reaction and Applications

• Item-derived application: Expression of an sgRNA designed to target CBLL2 using a lentiviral (pLenti) vector context. Suitable for research workflows involving CRISPR‑based perturbation where an external nuclease (e.g., Cas9 or Cas variants) is provided by co‑transfection, stable expression, or infection (general/literature context). No specific application conditions are provided in the Product Data.
• Typical research uses (literature/general, not item‑specific):
  • Gene knockout via CRISPR/Cas9 NHEJ at CBLL2.
  • CRISPR interference (CRISPRi) or activation (CRISPRa) when used with dCas9‑KRAB or dCas9‑VP64 systems, respectively, provided the spacer and promoter are compatible.
  • High‑throughput pooled or arrayed screening when integrated via lentiviral delivery (subject to IBC approval).
• Practical considerations (general, non‑procedural):
  • Confirm spacer sequence, on‑target locus, and PAM compatibility in silico before use; sequence‑verify any critical regions.
  • Validate on‑target editing/perturbation via orthogonal assays (e.g., amplicon sequencing, qPCR of target transcripts, or protein readouts), and assess potential off‑target effects appropriate to the study design.
  • Maintain appropriate controls (non‑targeting sgRNA, safe‑targeting sgRNA, and a known positive control), per good research practice. Note: No procedural steps, transfection conditions, or packaging recipes are provided in the Product Data; consult institutional protocols and the SDS/CoA for any item‑specific constraints.

Reaction Conditions

Not specified for this item; refer to CoA/Spec Sheet. As a general note (literature/general), conditions for successful use of sgRNA plasmids depend on the chosen delivery method (e.g., transfection, electroporation, or lentiviral transduction), the nuclease system employed, and the host cell type. Because procedural parameters are application‑ and institution‑specific, consult validated internal protocols and your biosafety committee for approved practices. No temperatures, timings, or reagent ratios are provided in the Product Data.

Safety and Handling

• GHS classification, signal word, pictograms, H‑statements — Not specified for this item; refer to SDS.
• Biosafety considerations (general): Plasmid DNA encoding sgRNA is non‑replicating outside appropriate hosts but should be handled under institutional biosafety guidelines for recombinant DNA. If used to generate lentiviral particles, follow BSL‑2 or higher practices as mandated by your IBC.
• PPE (general best practice): Lab coat, gloves, and eye protection. Use nuclease‑free technique to avoid cross‑contamination.
• Incompatibilities (general): Avoid nucleases (DNase contamination), strong acids/bases, oxidants, and prolonged exposure to elevated temperatures. Do not autoclave formulated DNA.
• First aid (overview): In case of accidental contact, rinse with water; if inhaled as aerosol or if ingestion occurs, seek medical attention per institutional policy. Refer to SDS for authoritative guidance.
• Waste: Dispose of recombinant DNA materials in accordance with local regulations and institutional biosafety procedures (e.g., chemical disinfectants appropriate for surfaces; do not rely on bleach for DNA integrity if downstream analysis is planned).
• Transport: Shipped on dry ice packs + cold packs (item-specific). Maintain cold chain to preserve integrity. Always defer to the product SDS and your institutional biosafety committee for definitive safety requirements.

Solvent Selection

Not typically applicable. This product is a nucleic acid construct, generally supplied in an aqueous buffer or nuclease‑free water. If buffer composition is relevant to your application (e.g., salt sensitivity in downstream enzymology or transfection), consult the CoA/Spec Sheet. Avoid organic solvents which can denature or precipitate DNA (literature/general).

Storage and Reconstitution

• Storage conditions (item-specific): Store at -80°C.
• Shipping (item-specific): Shipped on dry ice packs + cold packs. Maintain cold chain upon receipt.
• Format, concentration, buffer, and reconstitution details — Not specified for this item; refer to CoA/Spec Sheet.
• General handling (non‑procedural):
  • Protect from repeated freeze–thaw by preparing working aliquots upon first use, using nuclease‑free materials (literature/general).
  • Avoid prolonged exposure to ambient temperatures; keep on cold blocks/ice during handling as appropriate.
• Stability: No shelf‑life or stability data are provided; verify integrity empirically (e.g., by agarose gel or spectrophotometry) prior to critical experiments (general guidance).

Structure and Identity

• Item type: Lentiviral sgRNA expression construct targeting the human gene CBLL2 (inferred from product name: pLenti-CBLL2-sgRNA).
• Item-specific identifiers: CAS, CID, InChIKey, SMILES — Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula and exact molecular weight — Not specified for this item; refer to CoA/Spec Sheet.
• Structural description (general, literature):
  • Typical pLenti sgRNA vectors are circular double‑stranded DNA plasmids bearing lentiviral long terminal repeats (LTRs), a packaging signal (Ψ), an sgRNA expression cassette (often under a U6 or H1 promoter), and a tracrRNA/scaffold sequence compatible with Cas nucleases. Selectable markers (e.g., antibiotic resistance) and reporters may be present depending on design. These features enable transcription of a single guide RNA that directs a co‑expressed or co‑delivered nuclease to the CBLL2 locus.
  • The 2D “structure” is best represented as annotated plasmid maps (restriction sites, cassettes). No small‑molecule stereochemistry applies.
• Sequence information: Not specified for this item; refer to CoA/Spec Sheet.
• Gene target (item-derived): CBLL2 (Cbl proto‑oncogene like 2). No spacer sequence or PAM context is provided in the Product Data.

Synthetic Utility

Not applicable in the classical synthetic chemistry sense. This product is a biological DNA construct rather than a small‑molecule building block or reagent. Its “utility” pertains to genetic perturbation studies (see Reaction & Applications and Biological Roles) rather than to chemical synthesis.

Target Specificity

• Antigen/epitope/clone/isotype — Not applicable; this is not an antibody despite the catalog category path label.
• Item-derived targeting: sgRNA construct denoted for CBLL2 by product name. Spacer sequence, PAM, strand orientation, and any off‑target assessment are Not specified for this item; refer to CoA/Spec Sheet.
• Species context (genome build/isoform) — Not specified for this item; refer to CoA/Spec Sheet.