2d gel electrophoresis system

Introduction

A two dimensional (2D) gel electrophoresis system is a laboratory system used to separate the proteins and the other biomolecules by two properties, typically the isoelectric point in the first dimension and the molecular weight in the second dimension, and it is used in the proteomics, the biochemistry, and the research laboratories for the analysis of the complex protein samples. The 2D gel electrophoresis system consists of an isoelectric focusing (IEF) unit for the first dimension, in which the proteins are separated by the isoelectric point on the immobilized pH gradient (IPG) strips; a second dimension unit, in which the proteins are separated by the molecular weight by the SDS-PAGE on the polyacrylamide gel; and the power supply, the staining and the imaging equipment; the proteins are separated in the first dimension, transferred to the second dimension gel, separated by the molecular weight, and the gel is stained and imaged, and the resulting two-dimensional pattern of the protein spots provides a high-resolution map of the protein sample, which is analyzed with the image analysis software; the 2D gel electrophoresis system is used for the separation and the analysis of the complex protein mixtures, for the comparison of the protein expression between the samples, for the identification of the proteins, for the detection of the post-translational modifications, and for many other applications, and it is an essential technique in the proteomics. 2D gel electrophoresis systems are used in the proteomics research, in the biochemistry laboratories, in the clinical research, in the pharmaceutical industry, and for many other applications. With their high-resolution separation, 2D gel electrophoresis systems are essential for the protein analysis.

2D Gel Electrophoresis System LDGES-T2010

2D Gel Electrophoresis System first-dimension tube gel electrophoresis system features 120 mm and is compatible with various sample sizes. It has an advanced gel imaging system, providin...

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Standards

Complies with the above standards, but is not limited to them.

Features

1. 2D gel electrophoresis system for separating proteins by isoelectric point and molecular weight.

2. Includes isoelectric focusing (IEF) unit, second-dimension SDS-PAGE tank, power supply and cooling.

3. Suitable for proteomics, biomarker discovery, protein expression profiling and differential expression analysis.

4. Supports 2D gel electrophoresis (first dimension IEF, second dimension SDS-PAGE) with reproducible results.

5. Used in life science research, clinical proteomics, pharmaceutical R&D and agricultural biotechnology.

FAQ

Q1: What testing scenarios is this system mainly applied to?

A: The 2D gel electrophoresis system is used in proteomics research, biomarker discovery, protein expression profiling, and differential expression analysis to separate complex protein mixtures into individual protein spots based on two independent properties: isoelectric point (pI) in the first dimension (isoelectric focusing, IEF) and molecular weight in the second dimension (SDS-PAGE), enabling visualization and comparison of hundreds to thousands of proteins on a single gel. It supports proteomics research (global protein expression profiling of cells, tissues, biofluids (serum, plasma, urine, CSF); differential expression between control and treated/diseased samples; protein identification via spot excision and mass spectrometry (MALDI-TOF, LC-MS/MS); post-translational modification analysis (phosphorylation, glycosylation - shifts in pI/MW)), biomarker discovery (disease biomarkers (cancer, neurodegenerative, cardiovascular) - compare patient vs healthy; drug target identification; toxicity biomarkers (liver, kidney toxicity in preclinical studies)), pharmaceutical R&D (drug mechanism of action (protein expression changes after drug treatment), quality control of biopharmaceuticals (protein aggregates, degradation products, host cell proteins), batch consistency of protein therapeutics), clinical proteomics (disease subtype classification, treatment response monitoring, minimal residual disease), agricultural biotechnology (crop proteomics (stress response, GMO vs non-GMO), livestock disease markers, food authenticity (protein profiling for species identification, adulteration)), and microbiology (bacterial/viral proteomics, antibiotic resistance mechanisms, host-pathogen interactions). 2D electrophoresis principle: First dimension (IEF): proteins are separated in a pH gradient (immobilized pH gradient, IPG strip, typically pH 3-10 or 4-7) by applying an electric field; each protein migrates to its isoelectric point (where net charge = 0). Second dimension (SDS-PAGE): the IPG strip is equilibrated with SDS (denatures proteins, coats with negative charge proportional to length) and placed on top of a polyacrylamide gel; proteins separate by molecular weight (smaller migrate faster/farther). Result: 2D map with pI on x-axis, MW on y-axis. Proteins appear as spots; spot intensity correlates with abundance. Staining: Coomassie (simple, ~100 ng sensitivity), silver stain (~1 ng, more sensitive but tricky), fluorescent (SYPRO Ruby, DIGE - differential gel electrophoresis with CyDyes for accurate quantification). Compare to 3249 (pulsed field gel electrophoresis - for large DNA molecules) and 3248 (gel imaging system - for visualizing gels). This system is for protein 2D gels.

Q2: What standards does the system comply with?

A: The 2D gel electrophoresis system follows relevant ISO, ASTM, ICH and GB/T standards including ISO 15989 (proteomics - vocabulary, sample handling), ICH Q6B (biotechnological/biological products - test procedures and acceptance criteria for proteins, including electrophoresis), USP <726> (electrophoresis - for protein analysis in pharmacopeia), ASTM E1344 (protein electrophoresis), and GB/T standards for proteomics and electrophoresis. The hardware complies with applicable electrical safety (IEC 61010), EMC (IEC 61326), and performance (IEF voltage up to 10,000 V, temperature control at 20 C, SDS-PAGE cooling, power supply stability). For biopharmaceutical QC, follow ICH Q6B, USP <726>, and cGMP (21 CFR Part 211) - electrophoresis is used for purity, identity, and heterogeneity testing of protein therapeutics. For proteomics research, follow MIAPE (Minimum Information About a Proteomics Experiment) guidelines for data reporting and reproducibility. For regulated labs, maintain calibration records (power supply voltage/current, temperature, timer), equipment qualification (IQ/OQ), and SOPs. For gel documentation, use calibrated imaging system (3248). For DIGE (differential gel electrophoresis), follow manufacturer protocols for CyDye labeling and internal standard. Reagents (IPG strips, acrylamide, SDS, stains) should be analytical grade and from reputable suppliers.

Q3: What is the warranty period for the whole system and key components?

A: The whole 2D gel electrophoresis system is covered by a 12-24 month warranty (lab instruments often 1-2 years). The IEF unit, SDS-PAGE tank, power supply, cooling circulator, and controller are covered, while IPG strips (consumable - single use, expire; store at -20 C), acrylamide/bis-acrylamide (consumable - toxic, prepare or precast gels), SDS/buffers (consumable), stains (Coomassie, silver, SYPRO - consumable), and cooling fluid (wear - replace periodically) are consumables/accessories. Damage from improper use, high voltage safety issues (IEF uses up to 10 kV - ensure proper connections, no leaks), buffer leakage (corrodes electrodes - clean immediately), cooling failure (overheating causes IEF artifacts, gel smiling - check chiller), using expired IPG strips (pH gradient degradation - poor resolution), power surges (use UPS), or unauthorized repair is excluded. Extended warranty and service contracts (preventive maintenance, calibration, cooling system service) may be available. IEF unit 5-10 years; power supply 5-10 years; cooling circulator 3-7 years (pump wear); SDS-PAGE tank 5-10 years (glass plates break - replace). Consumables are ongoing significant cost (IPG strips $10-$30 each, precast gels $20-$50, stains).

Q4: Is this device difficult to operate? Is professional training required?

A: Operation requires trained molecular biology/proteomics personnel (familiar with electrophoresis, protein sample prep, IEF, SDS-PAGE). 2D electrophoresis is technically demanding (many steps, many artifacts) - professional training is strongly recommended. We provide operation manuals, installation, and training (2-5 days, including sample prep, IEF, SDS-PAGE, staining, imaging, analysis). Proper workflow (sample preparation (critical for 2D - remove salts, lipids, nucleic acids, detergents that interfere with IEF; use TCA/acetone precipitation or commercial cleanup kits; quantify protein (Bradford, BCA, 2D Quant); typical load 50-500 ug protein per IPG strip depending on stain), first dimension IEF (rehydrate IPG strip with sample (rehydration buffer: urea, thiourea, CHAPS, DTT, ampholytes), place in IEF tray, apply mineral oil to prevent evaporation/crystallization, run IEF program (stepwise voltage: e.g., 300 V 1 h, 1000 V 1 h, gradient to 8000 V, hold 8000 V until 40,000-80,000 Vh; total 12-24 h; maintain 20 C with cooling), equilibration (after IEF, equilibrate IPG strip in equilibration buffer with DTT (reduce disulfides) then iodoacetamide (alkylate, prevents re-oxidation); 15 min each; prepares proteins for SDS-PAGE), second dimension SDS-PAGE (place equilibrated IPG strip on top of SDS-PAGE gel (12.5% or gradient 8-16%), seal with agarose, run at constant current or voltage (e.g., 15 mA/gel 30 min then 30 mA/gel until dye front reaches bottom; 4-6 h; with cooling to prevent smiling/heating artifacts), staining (Coomassie (simple, MS-compatible, ~100 ng), silver (sensitive ~1 ng but glutaraldehyde-free for MS), SYPRO Ruby (fluorescent, sensitive, broad dynamic range, MS-compatible), DIGE (CyDye pre-labeling for quantitative comparison)), imaging (use gel imaging system (3248) with appropriate light source (white for Coomassie/silver, UV/blue for fluorescent), scan at resolution), analysis (2D gel analysis software (PDQuest, Melanie, SameSpots, ImageMaster) - spot detection, matching, quantification, differential expression (t-test, ANOVA), spot picking for MS identification), maintenance (daily: clean IEF tray/SDS-PAGE tank, check cooling fluid, wipe exterior; weekly: deep clean electrodes, check chiller level, inspect cables; monthly: calibrate power supply (voltage/current), verify temperature, clean cooling system; semi-annually/annually: professional service (high-voltage check, cooling pump, calibration)), troubleshooting (no spots / very few (protein load too low (increase), sample degradation (use protease inhibitors, keep cold), poor solubilization (increase chaotrope/detergent, use 2D cleanup), IEF failure (check voltage program, strip, connections), staining issue (optimize), horizontal streaking (poor IEF (salts, ionic detergents, insufficient Vh, IPG strip expired), protein overload (reduce load), poor sample prep (clean up), carrier ampholyte issue), vertical streaking (poor equilibration (increase time/reagents), SDS-PAGE issue (agarose seal, buffer), protein overload), smiling/frowning (overheating (check cooling, reduce current), gel polymerization issue, buffer imbalance), poor reproducibility (standardize sample prep, use same reagent lots, run technical replicates, use DIGE with internal standard for best quantification), and safety (HIGH VOLTAGE (IEF uses up to 10,000 V - electrocution hazard; ensure tray dry, no buffer spills, use manufacturer leads, do not touch during run; power supply has current limiting but still dangerous; SDS-PAGE uses 100-300 V - also hazardous), TOXIC REAGENTS (acrylamide is neurotoxic (wear gloves, mask when weighing; use precast gels to avoid; polymerized acrylamide is less toxic but still handle), silver stain uses formaldehyde, glutaraldehyde (toxic, carcinogenic - fume hood), SDS (irritant), DTT/iodoacetamide (toxic, wear PPE), Coomassie (methanol/acetic acid - flammable/corrosive), dispose as hazardous waste), COOLING (circulator contains water/antifreeze - do not spill on electronics; maintain level; use distilled water to prevent mineral buildup), do not use metal tools near high-voltage IEF tray, do not leave IEF/SDS-PAGE unattended for long (check for leaks, overheating), for MS identification: use MS-compatible stains (Coomassie, SYPRO Ruby; glutaraldehyde-free silver), excise spots with clean pipette tips, follow in-gel digestion protocol, for DIGE: use minimal labeling (Cy2 internal standard, Cy3/Cy5 samples), run on same gel for accurate comparison, scan with appropriate laser/filter, for reproducibility: use fresh reagents, standardized protocols, randomized sample order, technical/biological replicates, report per MIAPE guidelines, dispose of gels/reagents per hazardous waste regulations (acrylamide, silver, formaldehyde) are important for 2D electrophoresis.

Q5: What is the typical lead-time for order delivery?

A: Standard models require 7-30 working days. In-stock units can ship within 3-7 working days. Custom configurations or large built-to-order systems may require additional time, and we confirm the exact schedule before order confirmation.

Q6: Is regular calibration required? What is the recommended calibration cycle?

A: Calibration recommended every 6-12 months: power supply (voltage/current accuracy with multimeter, at multiple setpoints), temperature (IEF/SDS-PAGE cooling with calibrated thermometer at 20 C), and timer (with stopwatch). Routine checks: cooling temperature (before each run, verify 20 C), power supply output (periodic), and electrode condition (weekly - clean, no corrosion). If instrument is moved, cooling serviced, or results are inconsistent, recalibrate. For GMP/regulated labs (biopharma QC), follow applicable schedules and maintain records (calibration certificates, as-found/as-left data, thermometer ID, maintenance logs, IQ/OQ). For research labs, calibration is less formal but reproducibility requires consistent equipment performance. IPG strips and reagents are not calibrated but must be within expiration and stored properly (-20 C for strips). For DIGE, the imaging system (3248) requires separate calibration (fluorescence intensity, wavelength). 2D electrophoresis is more dependent on technique/reagents than instrument calibration - standardization of SOPs is critical for reproducibility.

Q7: Are supporting consumables universal?

A: IPG strips (specific pH range/length (7 cm, 11 cm, 13 cm, 18 cm, 24 cm; pH 3-10, 4-7, 5-8, etc.) - from Cytiva (Immobiline), Bio-Rad (ReadyStrip), or compatible; not universal across manufacturers (strip holder/tray size differs)), precast SDS-PAGE gels (specific to tank size - use OEM or compatible; hand-cast is alternative), reagents (acrylamide, SDS, buffers, stains - universal from Sigma/Bio-Rad/Thermo/Fisher), and cooling fluid (distilled water + antifreeze - universal) are available from life science supply companies, though IPG strips and some gels are system-specific (use OEM or certified compatible to fit IEF tray/SDS-PAGE tank). Long-term consumable cost is significant: IPG strips ($10-$30 each, single-use), precast gels ($20-$50), stains ($50-$200), buffers/reagents ($100-$500/month for active lab). DIGE CyDyes are expensive ($500-$1000 per set).

Q8: Can I customize parameters or supporting accessories?

A: Customization includes system configuration (IEF unit (single strip, 6-strip, 12-strip), SDS-PAGE tank (1 gel, 4 gels, 12 gels for high throughput), power supply (single output, multi-channel, programmable with stored methods), cooling (integrated or external circulator)), detection method (Coomassie/silver (standard, visible imaging), fluorescent (SYPRO Ruby, needs UV/blue imager), DIGE (Cy2/Cy3/Cy5 differential labeling, needs fluorescent imager with lasers, most quantitative)), automation (spot picking robot (automated excision for MS), gel casting robot, automated staining), imaging (integrated or separate gel documentation system (3248) with white/UV/fluorescent sources, high resolution CCD/camera), software (2D analysis software (PDQuest, SameSpots, Melanie, ImageMaster) for spot detection, matching, quantification, statistics, differential expression; MS identification integration), accessories (IPG strips (various pH/length), precast gels (various %/gradient), reagent kits (sample cleanup, equilibration, staining), CyDyes (for DIGE), molecular weight/pI markers, glass plates, casting stand, spot picking tools, cooling fluid, certifications (CE, ISO 9001, IEC 61010, ICH/USP compliance support), language and accessories. Additional consumables and accessories can be ordered separately.

Q9: Does the purchase include on-site installation and commissioning service?

A: Standard delivery includes 2D electrophoresis system (IEF unit, SDS-PAGE tank, power supply, cooling), and instruction manual (reagents/IPG strips often NOT included - order separately). On-site installation, setup, and operator training ARE typically included or strongly recommended for 2D systems (requires professional setup (cooling connection, power supply calibration, IEF tray installation), and user training on the technically demanding workflow (sample prep, IEF programs, equilibration, SDS-PAGE, staining, imaging, analysis); installation by manufacturer or certified technician is standard, especially for DIGE/high-throughput systems). Always run test gels with standard protein sample (e.g., E. coli lysate, serum) to verify system performance before experimental samples.

Q10: What types of samples are compatible with this equipment?

A: The 2D gel electrophoresis system separates proteins from various biological samples including cell lysates (mammalian cells, bacteria, yeast), tissue homogenates (liver, brain, tumor, plant), biofluids (serum, plasma, urine, CSF, saliva - may need depletion of high-abundance proteins (albumin, IgG) for deeper coverage), protein extracts (subcellular fractions, membrane proteins (need special solubilization), immunoprecipitates), and purified protein mixtures. NOT compatible with: Samples with high salt (>50 mM NaCl interferes with IEF - desalt or use cleanup), high detergent (ionic detergents like SDS interfere with IEF - use non-ionic (CHAPS, Triton) or remove SDS), nucleic acids (can cause streaking - treat with DNase/RNase or remove), lipids (cause smearing - delipidate or use cleanup), very high-abundance proteins (serum albumin >50% - masks low-abundance proteins; use depletion columns (MARS, Seppro)), extremely acidic/basic proteins (pH <3 or >10 may need specialized IPG strips (pH 3-12, 6-11) or nonequilibrium pH gel electrophoresis (NEPHGE)), very hydrophobic membrane proteins (poor solubilization in IEF - use special buffers (thiourea, ASB-14), or alternative methods (LC-MS)), very small (<5 kDa) or very large (>200 kDa) proteins (may run off gel or not enter - adjust gel %, use gradient), and non-protein samples (DNA, RNA - use agarose/polyacrylamide gel electrophoresis, not 2D protein gel; see 3249 PFGE for large DNA). For quantitative comparison: use DIGE (internal standard) for best accuracy; for simple profiling, Coomassie/silver with technical replicates. Sample load: 50-100 ug for analytical gels (silver/fluorescent), 500-1000 ug for preparative gels (Coomassie, spot picking for MS). IMPORTANT: 2D gel electrophoresis system fundamentals: (1) What is it? Separates proteins in 2 dimensions: - 1st: Isoelectric focusing (IEF) - by isoelectric point (pI) in pH gradient. - 2nd: SDS-PAGE - by molecular weight. Result: 2D protein map (spots). (2) Workflow: - Sample prep (cleanup, quantify, 50-500 ug). - IEF (IPG strip, 10-80 kVh, 20 C, 12-24 h). - Equilibration (DTT then iodoacetamide, 15 min each). - SDS-PAGE (12.5% or gradient, 4-6 h, cooled). - Stain (Coomassie/silver/SYPRO/DIGE). - Image (gel doc system). - Analyze (spot detection, matching, quantification, stats). - Identify (excise, in-gel digest, MS). (3) Key parameters: - IPG pH range: 3-10 (broad), 4-7 (most proteins), 5-8, 6-11 (basic). - IPG length: 7 cm (quick), 11/13 cm (standard), 18/24 cm (high resolution). - IEF: up to 10,000 V, 40-80 kVh. - SDS-PAGE: 12.5% (standard), 8-16% gradient (broad MW). - Load: 50-100 ug (analytical), 500-1000 ug (preparative). (4) Detection: - Coomassie: ~100 ng, simple, MS-compatible. - Silver: ~1 ng, sensitive, tricky (glutaraldehyde-free for MS). - SYPRO Ruby: fluorescent, ~1 ng, broad dynamic range, MS-compatible. - DIGE: CyDyes, most quantitative, internal standard, needs fluorescent imager. (5) Common artifacts: - Horizontal streaks: salts, poor IEF, overload. - Vertical streaks: poor equilibration, overload. - Smiling: overheating (cooling). - No spots: poor sample prep, load too low, stain failure. (6) Reproducibility: - Standardize sample prep and SOPs. - Use same reagent lots. - Technical replicates. - DIGE (best for quantitative comparison). - MIAPE reporting. (7) vs LC-MS proteomics: - 2D gel: visual map, PTM isoforms, lower cost, but limited dynamic range, hard for membrane/hydrophobic proteins, labor-intensive. - LC-MS: higher throughput, deeper coverage, better for membrane proteins, but no visual map, more expensive equipment. - Often complementary. (8) Choosing: - Basic proteomics: IEF + SDS-PAGE + Coomassie/silver. - Quantitative/differential: DIGE + fluorescent imager. - High throughput: 12-gel tank, automated staining/picking. - Biopharma QC: Validated system, USP/ICH compliance, IQ/OQ. This is distinct from 3249 (PFGE - for large DNA) and 3248 (gel imager - visualization only).