Native Protein Marker (66-669 kDa), IC-8066

Product Introduction

This product is a ready-to-use native protein molecular weight standard composed of four high-purity natural proteins, covering a molecular weight range from 66 kDa to 669 kDa. The product is formulated in an optimized native loading buffer and can be directly loaded onto native polyacrylamide gels (Native-PAGE) without boiling, reducing agents, or additional sample buffer. Simply thaw and mix before use.

Native-PAGE is an electrophoresis technique that separates proteins while preserving their native conformation. Protein migration depends on molecular weight, charge density, and molecular shape. This product is specifically designed for native electrophoresis and produces clear, evenly distributed bands after Coomassie Blue staining, suitable for molecular weight estimation of target proteins under native conditions, protein complex analysis, and enzyme activity assays. This product is not suitable for denaturing electrophoresis (SDS-PAGE), as SDS causes subunit dissociation and abnormal band migration.

Product Features

1. Ready-to-Use: Pre-mixed in native loading buffer; thaw and load directly.

2. High Molecular Weight Coverage: Covers 66-669 kDa, suitable for large proteins and protein complexes.

3. Clear and Uniform Bands: Optimized protein ratios produce uniform staining intensity after Coomassie Blue staining.

4. Good Stability: Stable for 12 months at -20¡ãC with good tolerance to repeated freeze-thaw cycles.

5. Broad Compatibility: Compatible with standard Tris-Glycine native gel systems and various native electrophoresis buffers.

Specifications

Size: 200 µL

Band Sizes: 66, 140, 440, 669 kDa

Form: Ready-to-use liquid in native loading buffer

Application: Molecular weight reference for Native-PAGE

Storage and Stability

Storage Conditions: Store at -20¡ãC, protected from light. After first use,

aliquot into single-use volumes (5-10 µL) to avoid repeated freeze-thaw cycles.

Shelf Life: 12 months from date of manufacture in unopened vials.

Protocol (For Reference Only)

Important: Thaw at room temperature or on ice. Mix gently by inversion to dissolve any precipitate. Do not boil or heat.

1. Native Gel Electrophoresis:

1.1 Prepare native polyacrylamide gel at the desired concentration. Tris-Glycine system recommended: 6-10% separating gel, 4% stacking gel.

1.2 Assemble the electrophoresis apparatus and fill with 1¡Á native electrophoresis buffer (Tris-Glycine, pH 8.3 recommended).

1.3 Load 3-5 µL of this product per well. For a 0.75 mm ¡Á 5 mm well, 5 µL is recommended. Adjust for other well sizes.

1.4 Run at constant voltage: 80-100 V for stacking gel, 120-150 V for separating gel. Stop when bromophenol blue front is approximately 1 cm from the gel bottom.

2. Staining and Visualization:

2.1 After electrophoresis, stain the gel with Coomassie Blue staining solution at room temperature for 1-2 hours.

2.2 Destain until background is clear and bands are distinct.

2.3 Document using a gel imaging system. For silver staining, dilute the Marker 50-fold before loading to avoid signal oversaturation.

3. Molecular Weight Estimation:

Reference molecular weights: 669 kDa, 440 kDa, 140 kDa, 66 kDa.

Precautions

1. Not for SDS-PAGE: This product is designed for Native-PAGE only. SDS will disrupt native conformation and cause abnormal band migration.

2. Do Not Heat: Thaw and mix only. Do not boil or heat.

3. Prevent Protease Degradation: Use sterile tips. Close the tube immediately after use.

4. Aliquot Storage Recommended: Aliquot into 5-10 µL per tube after first use. Use each aliquot once.

5. Molecular Weight Estimation Caution: In native electrophoresis, protein migration is affected by molecular weight, charge, and shape. Single gel concentration cannot provide precise absolute molecular weight. For precise determination, measure Rf values at multiple gel concentrations and construct a Ferguson plot.

6. Recommended Electrophoresis System: Tris-Glycine buffer (pH 8.3-8.8) with 8% separating gel provides ideal band spacing. Other buffer systems or gel concentrations may cause band shift.

FAQ (Simplified)

Q1: Bands are blurry or show tailing. What should I do?

A1: Check the following: (1) Mix thoroughly by gentle inversion 5-10 times after thawing. (2) Excessive freeze-thaw cycles may cause protein degradation; use a fresh aliquot. (3) Buffer pH is outside recommended range; prepare fresh buffer. (4) Gel polymerization is uneven; prepare a new gel.

Q2: Fewer bands than expected. What could be the cause?

A2: (1) Confirm the product was not used for SDS-PAGE. (2) Increase loading volume to 5-8 µL. (3) Extend staining time or use fresh staining solution.

Q3: Can this product be used for Western Blot?

A3: Not recommended. Transfer efficiency of native proteins is variable, and different proteins bind membranes differently, causing incomplete or uneven signal. Use prestained protein markers for Western Blot.

Q4: What if my target protein is outside this molecular weight range?

A4: This product covers 66-669 kDa. For proteins below 66 kDa, use a low molecular weight native marker. For proteins above 669 kDa, use an ultra-high molecular weight marker or estimate by Rf extrapolation.

Q5: The 440 kDa and 669 kDa bands are too close to distinguish. How can I improve resolution?

A5: (1) Reduce separating gel concentration to 6%. (2) Extend electrophoresis time. (3) Reduce voltage to 100-120 V.

Q6: Marker bands are too intense with silver staining and obscure sample signals.

A6: Dilute this product 50-fold before loading for silver staining, or load the Marker in a separate lane.

Disclaimer

1. For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for use in food or drugs.

2. Due to the exploratory nature of scientific research, conduct pilot experiments before applying to valuable samples.

3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages.

4. Wear appropriate protective clothing, gloves, and mask when handling this product.

Ordering Information

Catalog Number: IC-8066

Product Name: Native Protein Marker (66-669 kDa)

Size: 200 µL

Suggested Retail Price: CNY ¥1200.00 / USD $120.00 / EUR €132.00 / JPY ¥25956.00

Note: The specific price is subject to the transaction price in the offline contract. This document is provided for technical reference only and does not constitute any form of sales advertisement or invitation to offer. For product details and purchase inquiries, please contact local sales representatives.