EMSA Probe STAT5 - 1.75μM

Cat. No.: E745619

グレード＆純度 1.75μM

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

概要

Aladdin's EMSA Probe – STAT5 is a STAT5 consensus oligonucleotide designed for EMSA . This double-stranded oligonucleotide contains a well-recognized site for STAT5 transcription factors, and can be used as a probe for EMSA. The nucleotide sequence of STAT5 consensus binding oligo is as follows

Precautions：

Avoid heating above 40℃, as double-stranded DNA probe will be denatured at high temperatures into single strands that cannot be used in EMSA.For detailed EMSA procedures, please refer to the manual of EMSA kits from .This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1.Set up the probe labeling reaction with isotopes by adding reagents in sequence as follows:Probe to be labeled (1.75µM)2µlT4 Polynucleotide Kinase 10X Buffer1µlNuclease-Free Water5µl[γ-32P]ATP (3,000Ci/mmol at 10mCi/ml)1µlT4 Polynucleotide Kinase (5–10u/µl)1µlTotal volume10µl2.To label the probe with biotin, please referred to the EMSA Probe Biotin Labeling Kit (, GS008).3.For cold competition reaction setup, the amount of unlabeled probe should be 50-100 times that of labeled probe.

仕様

仕様と純度

1.75μM

安定性と保管

Store at -20℃ for up to 1 year.

保管条件

Store at -20°C

入荷

Ice chest + Ice pads

この製品は冷冷この冷このこの製品の運送が必要です。地上およびその他の経済サービスは利用できません。

Product Line

生物製品

Application Protocols

Item-specific tested applications, dilutions, and positive controls:

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

Note:

• While EMSA is the implied application from the product name, no lot-validated protocol steps, probe concentration ranges, or detection method are provided for SKU E745619. Use the general EMSA guidance in the Reaction & Applications and Reaction Conditions sections as starting context, and verify details against your internal SOPs.

Biological Roles

General background on the biological target (literature; not item-specific):

• STAT5 refers to Signal Transducer and Activator of Transcription 5, typically the STAT5A and STAT5B paralogs. STAT5 proteins are latent cytoplasmic transcription factors activated via phosphorylation by JAK kinases following cytokine or growth factor receptor engagement.
• Upon phosphorylation, STAT5 dimerizes, translocates to the nucleus, and binds GAS-like DNA elements with a core consensus TTCNNNGAA to regulate transcription of target genes.
• EMSA probes containing STAT5 motifs enable in vitro detection of STAT5–DNA complex formation from cell or tissue extracts, aiding studies of JAK–STAT pathway activity, DNA-binding specificity, and effects of post-translational modifications or co-factors.
• Probe competition (wild-type vs. mutated motif) and supershift with STAT5-specific antibodies are standard controls to establish specificity.

Note for SKU E745619:

• Biological function of STAT5 summarized above is provided for context. The exact sequence, motif spacing, and labeling for this probe are Not specified for this item; refer to CoA/Spec Sheet for definitive details.

Buffer Applications

Relevance:

• This product is an EMSA probe, not a pH buffer. Nevertheless, binding and electrophoresis buffers are central to EMSA performance.

General EMSA buffer guidance (literature; not item-specific):

• Binding buffer (example framework): 10 mM Tris·Cl pH 7.5–8.0, 50–150 mM KCl/NaCl, 0.5–1 mM DTT, 1 mM EDTA (or MgCl2 depending on protein), 5–10% glycerol, 50–200 µg/mL poly(dI·dC) as nonspecific competitor. Adjust salt and divalent ions to the protein’s optimal conditions.
• Electrophoresis buffer: 0.5× TBE is commonly used for native PAGE (4–6% polyacrylamide for 20–30 bp probes). Run at 4–10 °C to preserve complexes.
• Sample buffer: add a small amount of bromophenol blue/xylene cyanol without SDS; maintain glycerol for density.
• Post-electrophoresis handling: for non-radioactive probes, transfer to nylon membrane and perform chemiluminescent detection as appropriate for the label.

For SKU E745619, no item-specific buffer recipes or kit components are specified. Consult the CoA/Spec Sheet for any recommended buffer compositions tailored to this probe.

Green Alternatives

Context:

• EMSA probes historically used radioisotopic labeling (e.g., 32P). Modern non-radioactive labels reduce environmental, safety, and regulatory burdens.

General comparison (literature; not item-specific):

• 32P radiolabel vs. biotin/DIG/fluor labels:
  • Hazards/Waste: radioisotopes require specialized handling and waste disposal; non-radioactive labels avoid radiological hazards.
  • Sensitivity: 32P remains exceptionally sensitive; biotin–streptavidin chemiluminescence and DIG–anti-DIG HRP can approach comparable sensitivity with optimized protocols.
  • Workflow: non-radioactive methods offer longer signal stability, no decay, and simpler logistics.
  • Equipment: radioisotopes need shielding and film/PhosphorImager; non-radioactive detection uses standard blot imagers or chemiluminescence.

Sustainable practices:

• Use non-radioactive detection when experimental sensitivity permits.
• Prepare only required aliquots to minimize waste; store at −20 °C to extend shelf life and reduce remakes.
• Choose aqueous buffers and avoid unnecessary organic solvents.

Note: The label chemistry for SKU E745619 is Not specified for this item; refer to CoA/Spec Sheet for confirmation.

Pharmaceutical Uses

Applicability:

• EMSA probes are research tools for analyzing protein–DNA interactions. They are not pharmaceutical ingredients or excipients.

Relevant non-clinical contexts (general; not item-specific):

• Process and discovery research: assessment of pathway activation in cell-based systems, characterization of recombinant proteins, and mechanism-of-action studies in preclinical R&D.
• QC/analytics in research settings: qualitative monitoring of DNA-binding activity during protein purification or assay development.

No pharmacopeial status, formulation role, or clinical use is indicated or implied for SKU E745619. For research use only (per Product Data).

Physical Properties

Item-specific specifications:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight, exact length (nt), extinction coefficient, and label: Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties of dsDNA EMSA probes (for context only):

• Physical state: supplied lyophilized or as an aqueous solution, depending on product; dissolves readily in nuclease-free water or TE buffer.
• Solubility: highly water-soluble; insoluble in most organic solvents.
• pKa/logP: not conventionally defined for oligomers; behavior dominated by polyanionic phosphate backbone.
• UV absorbance: peak near 260 nm; typical quantitation at A260 (1 A260 ≈ 33 µg/mL for ssDNA; ≈50 µg/mL for dsDNA; literature values vary with length/base composition).
• Tm (melting temperature): depends on sequence, length, salt, and concentration; STAT-element probes (≈18–30 bp) often exhibit Tm in the 55–75 °C range under standard EMSA salt conditions (literature, sequence-dependent).
• Storage stability: stable at −20 °C when dry or in buffer; repeated freeze–thaw should be minimized (general practice).

Do not treat the above as specifications for SKU E745619. For definitive values (e.g., concentration, label, length, and extinction coefficient), refer to the item’s CoA/Spec Sheet.

Quality and Grades

Item-specific grade/purity details:

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

Context for nucleic-acid probe quality (general; not item-specific):

• Purification: EMSA-grade DNA probes are commonly HPLC- or PAGE-purified to remove truncated species and free label. The exact purification method for this SKU is Not specified for this item.
• QC metrics: typical data include analytical HPLC/PAGE trace, OD260-based quantitation, calculated vs. measured mass (MALDI/ESI), and verification of duplex formation. Some vendors provide functional QC via a reference EMSA with a cognate protein; not specified here.
• Label quality: for non-radioactive probes (biotin/DIG/fluor), labeling efficiency and lot-to-lot consistency are critical. For radiolabeled probes, specific activity and incorporation yield are key. The label type and specifications for this SKU are Not specified for this item.
• Suitability: EMSA-grade implies low salt/organic contaminants that can affect binding or gel migration, and minimized nuclease contamination.

Recommendation:

• Consult the CoA/Spec Sheet for this lot’s purification method, concentration or mass delivered, labeling chemistry (if any), and recommended annealing/handling instructions.

Reaction and Applications

Relevant application family:

• Electrophoretic Mobility Shift Assay (EMSA) to monitor STAT5 DNA-binding activity in nuclear or whole-cell extracts and purified systems.

General/literature use cases (not item-specific):

• Binding assays: quantify STAT5–DNA complex formation, determine apparent Kd, and compare binding in response to pathway activation (e.g., cytokine-stimulated JAK–STAT signaling in cell extracts). Competitive EMSA with unlabeled wild-type vs. mutant oligos helps confirm motif specificity.
• Supershift assays: addition of a STAT5 antibody yields a supershifted complex, supporting factor identity.
• Protein titrations and kinetics: assess binding stoichiometry and relative affinity across STAT5A vs. STAT5B or mutant proteins.
• Method notes: keep probe duplexed and on ice; pre-anneal complementary strands if supplied as single strands. Include poly(dI·dC) or salmon sperm DNA to suppress nonspecific binding. Optimize salt (50–150 mM KCl/NaCl), divalent cations (0–2 mM MgCl2), reducing agent (e.g., 0.5–1 mM DTT), glycerol (5–10%), and temperature (room temp to 4 °C) to stabilize complexes.
• Detection: depends on label (biotin/DIG/fluorescence/radio). Label for this SKU is Not specified for this item; refer to CoA/Spec Sheet.

Item-specific manufacturer applications: not provided beyond the product name. See general guidance above and consult the CoA/Spec Sheet for any lot-validated application details.

Reaction Conditions

Because this SKU’s validated protocols are not provided, the following is general EMSA guidance from literature (not item-specific; adjust per your system):

• Probe preparation: If supplied as single strands, anneal complementary oligos by heating to 90–95 °C for 2–5 min in annealing buffer (10 mM Tris·Cl, 50 mM NaCl), then slowly cool to room temperature. Store on ice.
• Binding reaction: 10–20 µL final volume; 1× binding buffer (e.g., 10 mM Tris·Cl pH 7.5–8.0, 75 mM KCl, 1 mM DTT, 1 mM EDTA or 1 mM MgCl2 as needed, 5–10% glycerol), 50–200 µg/mL poly(dI·dC), protein (extract or purified STAT5), and probe at 10–100 pM for highly sensitive labels or 0.1–1 nM for less sensitive labels. Incubate 15–30 min at room temperature or 4 °C.
• Competition/supershift: add 50–200× unlabeled specific competitor or mutant control; for supershift, include anti-STAT5 antibody and preincubate per vendor guidance.
• Electrophoresis: native PAGE 4–6% (29:1 acrylamide:bis), 0.5× TBE, pre-run 15–30 min; load samples and run at 100–150 V, 4–10 °C, until dye front reaches ~3/4 gel.
• Detection: dependent on probe label (chemiluminescent membrane detection for biotin/DIG; fluorescence imaging for fluor labels; autoradiography/PhosphorImager for 32P). Label for SKU E745619 is Not specified for this item; refer to CoA/Spec Sheet.

Optimize salt, divalent ions, and temperature for your specific STAT5 preparation and buffer system.

Safety and Handling

Item-specific hazard information:

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.

General guidance for nucleic acid probes (for context only; defer to SDS):

• Expected hazards: non-volatile, low inherent chemical hazard for unlabeled DNA probes; labeled probes may introduce additional hazards (e.g., acridinium esters, fluorescent dyes, DIG, or radioisotopes). This SKU’s label type is Not specified for this item; refer to CoA/Spec Sheet and SDS.
• PPE: laboratory coat, nitrile gloves, and safety glasses. Use RNase/DNase-free technique to prevent degradation.
• Handling: work on clean, nuclease-free surfaces; use dedicated pipette tips and tubes. Avoid repeated freeze–thaw; aliquot upon first thaw if supplied in solution.
• Incompatibilities: avoid strong nucleases (DNase), extreme pH (<6 or >9) or high temperatures unless intentionally used for annealing/denaturation. Keep away from oxidizing agents and UV light if fluorescently labeled.
• First aid (general): in case of contact with skin or eyes, rinse with water. If inhaled (aerosols) or ingested, seek medical attention. Provide SDS to responders.
• Waste: dispose according to institutional biosafety/chemical protocols. If radioactivity is involved (not specified here), follow radiological waste procedures.

Always consult the official SDS for SKU E745619 for authoritative hazard and first-aid information.

Solvent Selection

Applicability to this product type:

• EMSA DNA probes are handled in aqueous systems; organic solvent selection is typically not relevant.

General guidance (literature; not item-specific):

• Primary solvents/buffers: nuclease-free water, 1× TE (10 mM Tris·Cl, 1 mM EDTA, pH 7.5–8.0), or EMSA binding buffers supplied by kit protocols.
• Rationale: Tris buffer maintains pH near neutrality; EDTA chelates divalent cations to suppress nuclease activity during storage. During binding reactions, specific salts (e.g., KCl/NaCl), divalent ions (e.g., Mg2+), DTT, glycerol, and poly(dI·dC) are often included per protein requirements.
• Avoid: alcohols and most organic solvents; they destabilize duplex DNA and protein–DNA interactions and can denature proteins used in EMSA.
• Reconstitution tip: if supplied lyophilized, reconstitute with nuclease-free water to a convenient stock (e.g., 10–100 µM), then dilute into binding buffer immediately before use. This is general guidance; for SKU E745619, specific instructions are Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

Item-specific storage and shipping (from Product Data):

• Storage: Store at −20 °C.
• Shipping: Ice chest + ice pads.

Item-specific reconstitution details:

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

General guidance for DNA EMSA probes (not item-specific):

• Upon receipt: if lyophilized, briefly centrifuge the vial before opening. Reconstitute in nuclease-free water or TE buffer to a convenient stock concentration (e.g., 10–100 µM), based on the delivered mass or OD (check CoA). If provided in solution, verify concentration and label compatibility.
• Aliquoting: prepare single-use aliquots to avoid repeated freeze–thaw. Store aliquots at −20 °C in low-adsorption tubes. Protect from light if dye-labeled.
• Stability: duplexed probes are typically stable for months at −20 °C. Avoid multiple thaw cycles; thaw on ice and keep cold during setup.
• Handling: use DNase-free plastics and tips. For annealing (if needed), heat-complementary strands and slow-cool as described under Reaction Conditions.

For definitive instructions (e.g., recommended solvent/buffer, final assay concentration, and any label-specific precautions) for SKU E745619, consult the CoA/Spec Sheet.

Structure and Identity

Item-specific facts from Product Data:

• Product name: EMSA Probe STAT5 (SKU: E745619)
• 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 / InChI / InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

General/literature description (for context; not item-specific):

• This product name indicates a nucleic-acid probe intended for electrophoretic mobility shift assay (EMSA) targeting STAT5 DNA-binding activity. Such probes are typically short, double-stranded oligonucleotides containing a STAT5 consensus recognition element.
• Common STAT5 consensus motif (literature): TTCNNNGAA (gamma-activated sequence-like element). Exact sequence, length, and chemical modifications (e.g., biotin, DIG, fluorescent, or radioisotope labels) vary by product and are not specified here.
• Structural features (typical for dsDNA probes): phosphodiester backbone, nucleobases (A, T, G, C), potentially with 5' or 3' labels and terminal phosphates/caps depending on design.
• 2D description (generic): two antiparallel nucleotide strands forming a duplex with Watson–Crick base pairing around the STAT5 motif; any reporter tag (if present) is appended at an end via a linker.

Important note:

• Sequence, labeling chemistry, and strand design for SKU E745619 are Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Not typically applicable:

• EMSA probes are analytical/biochemical reagents rather than synthetic building blocks. They are not commonly used to form covalent bonds in organic synthesis.

Related methodological utility (general; not item-specific):

• As molecular tools, they enable quantitation of transcription factor binding, validation of consensus sequences, and benchmarking binding inhibitors in vitro.

For synthetic chemistry applications, refer to nucleic acid modification reagents or labeled nucleotide building blocks rather than finalized EMSA probes.

Target Specificity

Item-specific target details (validated data):

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

General/literature context (not item-specific):

• Intended target: STAT5 family transcription factors (STAT5A/STAT5B) that recognize GAS-like DNA elements.
• Typical recognition motif: TTCNNNGAA, sometimes arranged as tandem sites or palindromic configurations depending on promoter context. Precise sequence, spacing, and flanking nucleotides strongly influence affinity and specificity.
• Suggested controls: competition with unlabeled wild-type vs. mutated motif; supershift with STAT5-specific antibody; parallel assays with related STAT proteins to evaluate selectivity.

For SKU E745619, the exact oligonucleotide sequence and any proprietary modifications determining specificity are Not specified for this item; consult the CoA/Spec Sheet.