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| Product Name | Tris-Glycine-SDS Running Buffer, 10x Concentrate, Molecular Biology Grade |
| Catalog No. | ATR-B-YJL-0197 |
| Description | 10x concentrated Tris-Glycine-SDS (TGS) running buffer for SDS-PAGE electrophoresis in Tris-glycine gel systems (Laemmli method). The buffer contains 250 mM Tris base, 1.92 M glycine, and 1% (w/v) sodium dodecyl sulfate (SDS) at pH 8.3-8.5 (not adjusted). Upon dilution to 1x working concentration (25 mM Tris, 192 mM glycine, 0.1% SDS, pH ~8.3), it provides the necessary ions for electrical conductivity, maintains pH in the stacking (pH 6.8) and resolving (pH 8.8) gel regions, and provides SDS in the cathode (upper) buffer for maintaining protein denaturation and negative charge coating during electrophoretic migration. Prepared with ultrapure water, 0.1 um sterile-filtered, and quality tested for consistent SDS-PAGE performance. |
| Intended Use | Electrophoresis running buffer for denaturing SDS-PAGE analysis of proteins using standard Laemmli Tris-glycine gel systems (homemade or precast gels) in vertical electrophoresis tanks (Bio-Rad Mini-PROTEAN, Invitrogen XCell SureLock, Hoefer, and other standard systems). Suitable for reducing and non-reducing SDS-PAGE, western blotting gel separation, and Coomassie/silver staining workflows. |
| Principle / Technology | In SDS-PAGE, the Tris-glycine-SDS buffer system creates a discontinuous buffer system: chloride ions (from gel buffer) serve as leading ions with high mobility, glycinate ions (from running buffer) serve as trailing ions with lower mobility, and proteins complexed with SDS migrate between these boundaries. The Tris buffer maintains the appropriate pH gradient between stacking (pH 6.8) and resolving (pH 8.8) gels. SDS coats proteins with uniform negative charge density (approximately 1.4 g SDS per g protein), enabling separation based primarily on molecular weight. |
| Detection Method | Dilute 10x TGS buffer 1:10 with ultrapure water (e.g., 100 mL 10x + 900 mL water to make 1 L 1x buffer). Mix thoroughly. Fill electrophoresis tank: inner (cathode) chamber must be completely filled with 1x buffer to cover the wells; outer (anode) chamber filled to recommended level. Run SDS-PAGE at constant voltage (100-200 V) or constant current (20-40 mA per gel) until dye front reaches gel bottom. 1x buffer can be reused 2-3 times for consistent results when stored at RT. |
| Sample Type | 1x working solution (after 1:10 dilution): used in electrophoresis tank for SDS-PAGE separation of protein samples prepared in Laemmli sample buffer. |
| Performance Range / Specifications | 1x working concentration: 25 mM Tris, 192 mM glycine, 0.1% SDS, pH 8.3 +/-0.2, conductivity 4.5-5.5 mS/cm at 25 C; buffer capacity: sufficient for 3-4 gel runs per 1 L (1x) before significant pH shift; no detectable DNase, RNase, or protease activity; sterile-filtered through 0.1 um membrane. |
| Sensitivity / LOD | Not applicable — this is an electrophoresis running buffer, not a detection reagent. |
| Specificity | Formulated specifically for Laemmli Tris-glycine SDS-PAGE; not suitable for Tris-Tricine, Bis-Tris, Tris-Acetate, or other gel systems which require different running buffers. |
| Reaction Conditions / Protocol | Electrophoresis conditions: 100-200 V constant voltage or 20-40 mA per gel (constant current); typical run time 45-90 min at 150 V for 1.0 mm mini-gels; buffer temperature increase during run typically <15 C at 150 V. |
| Components / Formulation | 10x TGS Buffer Concentrate (1 L bottle): 250 mM Tris base (30.3 g/L), 1.92 M glycine (144.1 g/L), 1% (w/v) SDS (10 g/L), in ultrapure water, 0.1 um filtered, pH 8.3-8.5 (not adjusted). No preservatives or antimicrobial agents added. |
| Storage Conditions | Store at RT (15-25 C); SDS may precipitate at temperatures below 15 C — warm to RT and mix before use; do not freeze. |
| Shelf Life | 24 months from date of manufacture at RT; after dilution to 1x: 1 month at RT, 3 months at 2-8 C. |
| Package Specifications | 500 mL (makes 5 L 1x buffer), 1 L (makes 10 L 1x buffer), 4 L (makes 40 L 1x buffer). |
| Product Form | Clear, colorless to pale yellow liquid; 10x concentrate; may form slight precipitate at cold temperatures (SDS) — fully soluble upon warming. |
| Quality Control | Each lot tested for: pH (8.3-8.5 at 25 C), conductivity (4.5-5.5 mS/cm at 25 C for 1x), SDS concentration (0.095-0.105% 1x), DNase/RNase negative (incubation with lambda DNA and rRNA), protease negative (azocasein assay), sterility (0.1 um filtered, <1 CFU/mL), SDS-PAGE performance test: sharp band resolution of protein ladder (10-250 kDa) with linear Rf vs log(MW) plot (R2 >0.98). |
| Key Features | 10x concentrate for convenience; standard Laemmli formulation; 0.1 um sterile-filtered; nuclease and protease free; consistent lot-to-lot performance; 24-month shelf life; SDS included for protein denaturation. |
| Purity | All components ACS grade or higher; water: ultrapure (18.2 MOhm-cm); DNase, RNase, protease not detected; endotoxin <0.05 EU/mL. |
| Concentration | 10x concentrate: 250 mM Tris, 1.92 M glycine, 1% SDS; 1x working: 25 mM Tris, 192 mM glycine, 0.1% SDS. |
| Activity / Unit Definition | Not applicable — buffer solution. |
| Molecular Weight | Tris base: 121.14 g/mol; glycine: 75.07 g/mol; SDS: 288.38 g/mol (average). |
| Source / Origin | Synthetic chemicals (Tris, glycine from chemical synthesis; SDS from plant-derived fatty alcohol sulfation); water: ultrapure deionized; no animal-derived components. |
| pH Range / Optimal pH | 10x pH 8.3-8.5 (not adjusted); 1x pH approximately 8.3; Tris effective buffering range pH 7.0-9.0. |
| Shipping Conditions | Ambient temperature. |
| Expiration Date / Stability | 24 months at RT; SDS precipitation at low temperatures is reversible; do not freeze (freeze-thaw may cause irreversible precipitation). |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic use. Not for human or animal drug use. |
| Compatibility | Compatible with standard Laemmli SDS-PAGE using homemade or commercial Tris-glycine gels (any percentage). Compatible with all vertical electrophoresis systems. Can be used as anode (lower) and cathode (upper) buffer. 1x buffer is compatible with downstream western blotting (tank, semi-dry, and dry transfer systems), Coomassie Brilliant Blue staining (R-250 and G-250/Colloidal), silver staining, and fluorescent gel staining (SYPRO Ruby, Krypton). For western transfer, the presence of 0.1% SDS in 1x buffer may affect transfer of some proteins — for optimal transfer, include 10-20% methanol in transfer buffer. |
| Recommended Buffer System | 10x Tris-Glycine-SDS: 250 mM Tris, 1.92 M glycine, 1% SDS, pH 8.3-8.5; 1x: 25 mM Tris, 192 mM glycine, 0.1% SDS, pH ~8.3. |
| Application Notes / Precautions | If SDS has precipitated (white crystalline solid at bottom of bottle), warm the 10x buffer to 37 C in a water bath and mix thoroughly until completely dissolved before diluting. Do not microwave 10x buffer. Always use freshly diluted 1x buffer or buffer stored for no more than 1 month at RT. Reusing 1x buffer: the cathode (inner) buffer should not be reused as it accumulates SDS-depleted glycine and chloride ions, resulting in pH changes. The anode (outer) buffer can be reused 2-3 times. For best western blot results, use fresh 1x buffer for each transfer. If running very large proteins (>250 kDa), consider using Tris-Acetate gels with Tris-Acetate-SDS running buffer, as the glycine ions in Tris-Glycine-SDS may compress the resolving range. |
| Batch-to-Batch Consistency | pH within 8.3-8.5 for every lot; conductivity within 4.5-5.5 mS/cm (1x); SDS-PAGE Rf vs log MW linearity R2 >0.98 for every lot. |
For research use only, not for clinical use.
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