![SMOBIO/[PM2610] ExcelBand™ Enhanced 3-color High Range Protein Marker (9-245 kDa), 250 μl x 2/9-245 kDa), 250 μl x 2</span>
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Description
The PM2610/PM2611 ExcelBand™ Enhanced 3-color High Range Protein Marker is a ready to use three-color protein standard with 12 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine buffer (9 to 235 kDa in BisTris (MOPS) buffer and 10-235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with two reference proteins carrying enhanced intensity corresponding to a green at 25 kDa and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The ExcelBand™ Enhanced 3-color High Range Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins. The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading.
Features
Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
Two reference bands — 75 kDa (red) and 25 kDa (green)
Contents
Approximately 0.2~0.6 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25°C), 2 % SDS, 3.6 M Urea, and 15 % (v/v) Glycerol).
Quality Control
Under suggested conditions, the PM2610/PM2611 Enhanced 3-color High Range Protein Marker resolves 12 major bands in 15% SDS-PAGE (Tris-Glycine buffer) and after Western blotting to nitrocellulose membrane.
Storage
4°C for 3 months -20°C for 24 months
Specification
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Manual
Manual_PM2610_ExcelBand™ Enhanced 3-color High Range Protein Marker
SDS
SDS_PM2610
Migration patterns and approximate MWs (kDa)
Why are there contrasting results in molecular weights after using different brands of protein markers?
A.Different proteins even with similar molecular weights would exhibit apparent disparity from the resulting SDS PAGE due to the difference in the composition of the protein’s amino acids (e.g. gelatin). The reason for the disparity is due to the amino acids composition that affects the binding of the protein and SDS. Therefore, we can say that protein marker is a handy tool to estimate molecular weight, but there is no absolute molecular weight standard.
B.While running SDS-PAGE, protein mobility can be affected by the composition of the buffer used, gel percentage, the voltage used, running time, as well as if there is a pre-run.
C.Another recommendation for high molecular weight proteins is to prolong the running time to clarify the relative location of bands.
Protein marker Retention Period: Mentioned -20°C and over 2 years. Is it available for 30 months or 36 months? Have you tested this period?
Yes, we have tested our PM2700. The results showed that the PM2700 is stable at -20℃ for at least two years. It has also shown strong performance for more than 36 months under our careful storage. However, we must only suggest a 2 year retention period for the following reasons: There may be a variation in the environment in storage, and improper use may lead to accumulated damage to the proteins and therefore reduce its retention period.
How many times of freezing and thawing are available for protein markers? If it uses 5 μL per load, would the total usage quantity be 50 times x 2 (250 μL x 2 tube)?
Yes, 100 uses (5 μL each time) can be expected if freezing and thawing are conducted carefully and properly at the appropriate temperature. Before each use, make sure the protein marker is thoroughly thawed.
Do you have data comparison for protein molecular weight’s precision with other protein markers?
Yes. Usually, pre-stained marker is written on “estimated molecular weight” for caution. It is known that the analysis of protein size by an SDS-PAGE is only for “estimation” because of the intrinsic variation of amino acid composition in all proteins including stained and non-stained ones. For example, a protein which is highly hydrophilic might show a particular higher position in the SDS-PAGE analysis when compared to a hydrophobic one. We did compare the migration patterns of SMOBIO’s Protein Markers with other brands, and we concluded that it was difficult to define “precision” due to the reasons mentioned above. Therefore, in the product description, we suggest our users to calibrate the MW against their interested proteins. Although it is impossible to define "precision" for molecular weight of proteins in SDS-PAGE, we did compare the migration pattern of pre-stained markers with unstained protein marker (Invitrogen MARK12) for calibration. It is concluded that the estimated molecular weight of SMOBIO’s pre-stained marker shows a curve matching well with that of unstained native proteins (MARK12), representing a good estimation of the MW of each pre-stained protein in the SDS-PAGE analysis.
Will SMOBIO’s Protein Markers/Ladder be washed out during Western blotting process?
SMOBIO’s Protein Markers/Ladder will be only slightly washed out during Western blotting process. However, the excess of Tween-20 (more than 0.2%) in washing buffer will affect SMOBIO’s Protein Markers/Ladder on the transfer membrane.
Here are suggestions for Western blotting process:1. Transfer SMOBIO’s Protein Markers/Ladder to membrane with transfer buffer containing 20% methanol to fix SMOBIO’s Protein Markers/Ladder on membrane. 2. Wash membrane with PBS or TBS containing less than 0.1% Tween-20.
Will SMOBIO’s Protein Markers/Ladder be affected by the stripping/deprobing process with the presence of β-Mercaptoethanol (β-ME)?
In normal circumstances, the presence of βME during the stripping/deprobing process will only slightly affect SMOBIO’s Protein Markers/Ladder. However, the presence of Tween-20 on PVDF membrane during the stripping/deprobing process has adverse effects on SMOBIO’s Protein Markers/Ladder.
Here are suggestions for Western stripping/deprobing process:
1. Wash the PVDF membrane in methanol for 5~10 minutes prior to the stripping/deprobing process to mitigate the adverse effect of Tween-20.2. Recommended stripping buffer (for 1 L): 15 g glycine, 1 g SDS, 10 mL Tween 20. Dissolve in 800 mL distilled water. Adjust pH to 2.2 Bring volume up to 1 L with distilled waterUnraveling the novel effects of aroma from small molecules in preventing hen egg white lysozyme amyloid fibril formation
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ExcelBand™ Protein Markers
Ready-to-use— premixed with a loading buffer for direct loading, no need to boil
Broad range— 310 kDa to 5 kDa
Pre-stained bands — for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membrane
Enhanced bands— for quick reference
YesBlot™ Western Marker I
Ready-to-use — no need of mixing or heating before sample loading
Direct visualization — 10 IgG-binding proteins for direct visualization on Western blots
Pre-stained bands — 4 pre-stained proteins for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membrane
Wide range — 10 clear bands from 15 to 200 kDa for size estimation
Quick reference — two enhanced bands (30 and 80 kDa)
Q-PAGE™ Precast Gels
User-friendly gel cassette:
Numbered and framed wells for sample loading
Labeled warning sign and green tape as reminder
Enhanced gel performance:
Enhanced gel electrophoresis speed
Better band separation
Stable for shipping at ambient temperature
Easy compatibility:
Available as homogeneous and adjusted gradient gels for a wide range of protein separation.
Compatible with most popular protein electrophoresis systems
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非常急！
蛋白质在水溶液中的溶解度是由蛋白质周围亲水基团与水形成水化膜的程度，以及蛋白质分子带有电荷的情况决定的。当用中性盐加入蛋白质溶液，中性盐对水分子的亲和力大于蛋白质，于是蛋白质分子周围的水化膜层减弱乃至消失。同时，中性盐加入蛋白质溶液后，由于离子强度发生改变，蛋白质表面电荷大量被中和，更加导致蛋白溶解度降低，使蛋白质分子之间聚集而沉淀。一般33%硫酸铵为沉淀Ig-G的最适饱和度
有机溶剂沉淀，极性有机溶剂降低介点常数使蛋白质凝集沉淀。eg.甲醇、乙醇、丙酮。
磺基水杨酸也可以沉淀蛋白质，但要加酸调节ph，使之低于血清蛋白的pi，使蛋白质带正电荷，与水杨酸酸根结合生成不溶盐而沉淀。
血清蛋白质均带负电荷，但各种蛋白质带负电荷的程度有所差异，以白蛋白为最多。而血清白蛋白等电点为4.7－4.9。所以把ph调到4.7以下就可以了。
