iBright™ Prestained Protein Ladder
iBright™ Prestained Protein Ladder
Invitrogen™

iBright™ Prestained Protein Ladder

iBright Prestained Protein Ladder contains twelve recombinant proteins, ten (11 to 250 kDa) that are blue-stained and fluor-labeled for directRead more
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Catalog NumberQuantity
LC56152 x 250 μL
LC5615X48 x 250 μL
Catalog number LC5615
Price (USD)
322.00
Each
Add to cart
Quantity:
2 x 250 μL
Price (USD)
322.00
Each
Add to cart
iBright Prestained Protein Ladder contains twelve recombinant proteins, ten (11 to 250 kDa) that are blue-stained and fluor-labeled for direct and near-IR fluorescent visualization and protein sizing, and two (30 kDa and 80 kDa) that are unstained and contain IgG binding sites to bind the primary and secondary antibodies used for chemiluminescent and fluorescent detection of the target protein. The protein ladder is supplied in a ready-to-use format for direct loading onto gels; no need to heat, reduce, or add sample buffer prior to use.

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Product features
• Fluorescent visualization—detect the 10 stained proteins using the 670 nm red laser or 700 nm channel of a fluorescence imager
• Western blot confirmation—detect the two unstained proteins (30 kDa and 80 kDa) using the detection system for the target protein

Applications
• Western blotting: detection of the two unstained bands via the detection method used for the target protein. Compatible with chemiluminescent substrates and fluorescent secondary antibodies.
• Protein sizing on SDS-polyacrylamide gels, visualization of proteins during electrophoresis and transfer, and visualization of proteins during fluorescence imaging (ten blue-stained bands)
• Protein size estimation: protein sizing on SDS-polyacrylamide gels and on western blot using near-infrared (NIR) fluorescence imagers

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodColorimetric, NIR Fluorescence, and User defined detection system (2 bands)
Number of Markers12
Product TypeProtein Ladder
Ready to LoadYes
Size Range11 to 250 kDa
Stain Type1 color: Blue
Gel CompatibilityNovex™ Midi Gels, Novex™ Mini Gels, Novex™Tris-Glycine Gels, NuPAGE™ Bis-Tris Gels, NuPAGE™ Gels, NuPAGE™ MES Gels, NuPAGE™ MOPS Gels, NuPAGE™ Tris-Acetate Gels, Precise™ Tris-Glycine Gels, SDS-PAGE Gels, Precise™ Tris-HEPES Gels
Molecular Weight (g/mol)250, 130, 95, 80, 70, 55, 43, 34, 30, 26, 15, 11 kDa
Product LineiBright™
Quantity2 x 250 μL
Shipping ConditionDry Ice
System TypeWestern Blotting, SDS-PAGE
Unit SizeEach
Contents & Storage
Storage buffer: 62.5 mM Tris-H3PO4 (pH 7.5 at 25°C), 1 mM EDTA, 2% (w/v) SDS, 10 mM DTT, 1 mM NaN3 and 33% glycerol

Upon receipt, store at –20°C. Product is shipped on ice.

Frequently asked questions (FAQs)

What gel running buffer should I use with the iBright Prestained Protein Ladder? Tris-glycine, Tris-tricine, or Tris-acetate running buffer?

Most of the common gel running buffers are composed of Tris-glycine or Tris-tricine. Tris-glycine buffer systems are useful for separation of proteins over a wide range of molecular weights (5-300 kDa) and are compatible with denaturing or non-denaturing conditions. Tris-tricine buffer is generally recommended for the electrophoresis of low molecular weight proteins and peptides (<10 kDa) that need to be reduced and denatured prior to loading. Tris-acetate buffer system is used for separation of larger proteins (>200 kDa).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can the iBright Prestained Protein Ladder be detected by IgM antibodies?

The 2 unstained bands in the iBright Prestained Ladder contain an IgG binding site, allowing direct visualization with the same antibody-conjugate reagents used to detect the target protein. The proteins in the standard will not bind to IgM antibodies.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

With the iBright Prestained Protein Ladder, I am unable to detect the 2 unstained protein bands. Why is this?

Primary antibodies with low starting concentrations may result in insufficient chemiluminescent detection of the western blot positive control bands. If the unstained 30 kDa and 80 kDa bands produce weak or no signal, spike the diluted primary antibody with the corresponding rabbit IgG or mouse IgG to a concentration of 1-5 µg/mL, prior to secondary antibody incubation. Follow with respective secondary (GAM/GAR) incubation to increase the intensity of western blot positive control bands in the iBright Prestained Protein Ladder.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What is the recommended gel loading volume for the iBright Prestained Protein Ladder?

If performing detection on a mini gel, the recommended loading volume is 1-3 µL. If performing visualization on a midi gel, 2-4 µL is recommended, and for detection, 2-3 µL is recommended. Optimal loading volume should be determined empirically.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

When should I use the iBright Prestained Protein Ladder over other protein ladders?

The iBright Prestained Protein Ladder has been optimized and designed for use with western blotting as it provides 2 control bands (unstained ladder bands) that are detected using the same antibody conjugate and protocol. The 2 unstained control bands contain IgG binding sites that can be visualized simultaneously with your target without any additional steps in the protocol. The 10 stained bands will appear during electrophoresis and transfer. These 10 bands will also appear in fluorescent imaging in the NIR channels. The 2 control bands will appear with chemiluminescent or fluorescent detection similar to your target.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.