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  6. Invitrogen™ NuPAGE™ LDS Sample Buffer (4X) 

Invitrogen™ NuPAGE™ LDS Sample Buffer (4X)

Product Code. 11549166
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Product Code. 11549166 Supplier Invitrogen™ Supplier No. NP0007

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Description

Includes

NuPAGE LDS Sample Buffer (4X), 10mL

Used to prepare protein samples for denaturing gel electrophoresis with NuPAGE™ Novex™ gels. Invitrogen™ Novex™ NuPAGE™ LDS Sample Buffer (4X) contains lithium dodecyl sulfate, pH 8.4, which allows for maximum activity of the reducing agent.

Optimal protein sample preparation for polyacrylamide gel electrophoresis (PAGE) requires denaturing and reducing protein disulfide bonds.

  • Highly viscous and concentrated solution that contains higher a concentration of glycerol
  • Contains twice the amount of LDS compared to the amount of SDS in Novex™ Tris-Glycine SDS Sample Buffer or in Tricine SDS Sample Buffer
  • NuPAGE™ LDS Sample Buffer should be brought to room temperature (25°C) prior to use

Order Info

Shipping Condition: Approved for shipment at Room Temperature or on Wet Ice

TRUSTED_SUSTAINABILITY

Specifications

Content And Storage Sample Buffer (4X) containing lithium dodecyl sulfate (LDS) at pH 8.5 with SERVA Blue G250 and phenol red.

Store at 4°C to 25°C.
Buffer Sample Loading Buffers
Concentration 4 X
Gel Type NuPAGE
Gel Compatibility NuPAGE™ Gels
Quantity 10mL
Shipping Condition Approved for shipment at Room Temperature or on Wet Ice
Product Line NuPAGE
Product Type LDS Sample Buffer

Frequently Asked Questions (FAQs)

Can I prepare my protein sample with the reducing agent and store it for future use?

DTT is not stable, so it must be added and the reduction performed just prior to loading your samples.
My LDS or SDS sample buffer precipitates when stored at 4 degrees C. Can I warm it up? Can I store it at room temperature?

Precipitation of the LDS or SDS at 4 degrees C is normal. Bring the buffer to room temperature and mix until the LDS/SDS goes into solution. If you do not want to wait for it to dissolve, you can store the sample buffer at room temperature.
How are Bolt gels different than NuPAGE gels?

While they are both Bis-Tris based gels, the chemistries are very different since Bolt gels are optimized for western blotting. Another key difference is the wedge well design of the Bolt gels, which allows larger sample volumes to be loaded.
What is the advantage of NuPAGE Gels over regular Tris-Glycine gels?

The neutral operating pH of the NuPAGE Gels and buffers provides following advantages over the Laemmli system:
-Longer shelf life of 8-12 months due to improved gel stability
-Improved protein stability during electrophoresis at neutral pH resulting in sharper band resolution and accurate results (Moos et al, 1998)
-Complete reduction of disulfides under mild heating conditions (70 degrees C for 10 min) and absence of cleavage of asp-pro bonds using the NuPAGE LDS Sample buffer (pH > 7.0 at 70 degrees C)
-Reduced state of the proteins maintained during electrophoresis and blotting of the proteins by the NuPAGE Antioxidant
Please refer to the following paper: Moos M Jr, Nguyen NY, Liu TY (1988) Reproducible High Yield Sequencing of Proteins Electrophoretically Separated and Transferred to an Inert Support. J Biol Chem 263:6005-6008.
What is the composition of the NuPAGE LDS Sample Buffer and what is the recipe for the 4X formulation?

The composition of the 1X NuPAGE LDS Sample Buffer is as follows:

141 mM Tris base
106 mM Tris HCl
2% LDS
10% Glycerol
0.51 mM EDTA
0.22 mM SERVA Blue G
0.175 mM Phenol Red
pH 8.5


To prepare 10 mL of 4 X NuPAGE LDS Sample Buffer, dissolve the following reagents to 8 mL ultrapure water:

Tris HCl 0.666 g
Tris Base 0.682 g
LDS 0.800 g
EDTA 0.006 g
Glycerol 4 g
SERVA Blue G (1 solution) 0.75 mL
Phenol Red (1 solution) 0.25 mL

Mix well and adjust the volume to 10 mL with ultrapure water. Store at +4. The buffer is stable for 6 months when stored at +4°C.
Can a protein sample be boiled in NuPAGE LDS Sample Buffer? Are there any components that may be destroyed by boiling?

You may boil the protein sample, although we recommend heating at 70°C for 10 min instead. The buffer components are not damaged during boiling. Heating rather than boiling will produce sharper protein bands, and this may be due to reduced protein hydrolysis at the lower temperature.
What is the pH of the NuPAGE LDS Sample Buffer (4X) at 70°C?

The temperature coefficient (ΔpKa/degree C) for Tris is -0.031/degree C. Therefore, if the pH of the NuPAGE LDS Sample Buffer is 8.5 at 25°C, then its pH at 70°C is 8.5 + (-0.031 X 45) = 7.1. Similarly, our Tris Glycine Sample Buffer at 85°C would have a pH of 4.94, at 100°C its pH would be 4.5.
Can beta-mercaptoethanol (BME) be used rather than DTT as the reducing agent in the NuPAGE LDS Sample Buffer?

Either BME or DTT can be used in the NuPAGE LDS Sample Buffer.

Make sure that a fresh solution of BME is used. FINAL concentration:

DTT 50-100 mM

BME 2-5%
What is the recipe for traditional Laemmli Buffer?

The Laemmli buffer or 2X SDS Buffer is composed of the following: 100 mM Tris HCl , pH 6.8, 200 mM dithiothreitol, 4% SDS, 0.2% bromophenol blue, 20% glycerol. 2X SDS gel loading buffer lacking dithiothreitol can be stored at room temperature. Dithiothreitol should then be added, just before use.
Why is LDS (lithium dodecyl sulfate) used in the 4X NuPAGE sample buffer instead of SDS?

SDS in a 4X sample buffer concentrate tends to precipitate from solution and to make the solution viscous and difficult to pipette. The LDS is much more soluble.
Can I use CTAB rather than SDS in my sample buffer?

No, CTAB will not work with any of our gels except for the NuPAGE Tris-Acetate gels. To use CTAB, you would need to use a running buffer of 50 mM acetic acid and 50 mM beta-alanine in equal concentrations. You would also need to switch the electrodes. Since CTAB is a cationic detergent, this would establish conditions for running a basic protein towards the anode (into the gel).
Are the NuPAGE buffers/components (i.e., sample buffer, running buffer, and antioxidant) compatible with Bolt gels?

Yes, NuPAGE buffers/components are compatible with Bolt gels.

I have prepared my protein sample using NuPAGE LDS Sample buffer. Can I boil my samples?

It would be okay to boil the protein sample as none of the buffer components are damaged upon boiling. The reason we recommend to heat the sample at 70 degrees C for 10 minutes and not boil the sample is because protein hydrolysis is reduced at lower temperatures.

Why is LDS used in the NuPAGE Sample buffer instead of SDS?

The pH of the NuPAGE 4X Sample buffer is much higher than that of the usual Laemmli Sample buffer: 8.5 vs. 6.8. At this pH (8.5), SDS will precipitate when added at the concentration needed for a 4X buffer. On the other hand, LDS (lithium dodecyl-sulfate) is more soluble and is just as effective for denaturation.
How should I store the NuPAGE buffers?

NuPAGE buffers can be stored at 4-25 degrees C.

For Research Use Only. Not for use in diagnostic procedures.

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