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Thermo Scientific™ Pierce™ Reversible Protein Stain Kit for PVDF Membranes

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PI24585 1.75 L
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Catalog No. PI24585 Supplier Thermo Scientific™ Supplier No. LSG24585
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Description

Reversibly stain proteins on nitrocellulose or PVDF membrane after transfer from polyacrylamide gels with this sensitive Ponceau S alternative.

Thermo Scientific™ Pierce Reversible Protein Stain Kits for Membranes are rapid and sensitive alternatives to Ponceau S stain for reversible detection of protein bands on nitrocellulose membrane or PVDF membrane after transfer from polyacrylamide gels.

These kits for membrane-staining use a nondestructive, reversible, reliable and sensitive method to stain and detect proteins on nitrocellulose and PVDF membranes. These Pierce Kits are superior alternatives to Ponceau S stains for evaluating the efficiency of protein transfer following SDS-PAGE and before immunoblotting. The lower limit of detection with this method is 25 to 50ng per band (at least five times more sensitive than traditional Ponceau S staining). The staining protocols are simple, quick and result in turquoise-blue bands that do not fade and are easily photographed for future reference. The stain can be easily reversed in less than 15 minutes. Subsequent Western blot detection is unaffected because the stain does not alter the protein and is completely removed. Separate kits for nitrocellulose and PVDF membranes use the same dye-binding mechanism but differ in the stain formulation and membrane-treatment components to optimize the speed and performance with each type of membrane.

Highlights:

  • Better than Ponceau S – more sensitive, easier to document, permanent until reversed (does not fade)

  • Sensitive – high-avidity, total-protein stain; lower limit of detection equals 25 to 50ng per band

  • Specific – detects only protein; does not bind or interact with other electrophoresis or sample components

  • Rapid – stain in less than 5 minutes (nitrocellulose kit) or 30 minutes (PVDF kit); completely erase and reverse staining in less than 15 minutes

  • Stable – components are stable at room temperature, saving refrigerator space and eliminating equilibration steps

Includes:

Reversible protein stain, destain and eraser solutions; PVDF kit also contains a sensitizer solution

Specifications

Content And Storage Sufficient For: 10 PVDF membranes (8 cm x 8 cm) from mini gels (SDS-PAGE)
• Sensitizer, 250 mL
• Reversible Stain, 250 mL
• Destain, 1 L
• Stain Eraser, 250 mL

Store at room temperature.
Description Pierce Reversible Protein Stain Kit for PVDF Membranes
Detection Location In-Blot Detection
Detection Method Colorimetric
Label or Dye Proprietary Mix
Quantity 1.75 L
Target Molecule Protein
Product Line Pierce
Product Type Reversible Protein Stain Kit

Frequently Asked Questions (FAQs)

Are SuperBlock Blocking Buffers compatible with the Pierce Reversible Protein Stain Kit for PVDF Membranes (Cat. No. 24585)?

The procedure for the Reversible Protein Stain Kit for PVDF Membranes (Cat. No. 24585) is performed before the membrane is blocked with the blocking buffer, and thus, has no effect on the subsequent blocking.
How can I increase the signal intensity when I am using SuperSignal West Femto Maximum Sensitivity Substrate?

The following are possible causes for weak signal intensity:
- Low level of protein transferred from the gel to the membrane. This is particularly true for high molecular weight proteins. Reversible Protein Stain Kits for Nitrocellulose and PVDF membranes (Cat. Nos. 24580 and 24585, respectively) may be used to stain the bands on the membrane to indicate transfer efficiency.
- Over-diluted primary and secondary antibody probes. We suggest using the recommended dilution for the antibody probes for each substrate. More sensitive substrates will require a more dilute antibody solution.
- Short substrate incubation time. For best results, incubate the membrane for 5-15 min in the SuperSignal substrate.
How can I increase the signal intensity when I am using SuperSignal West Dura Extended Duration Substrate?

The following are possible causes for weak signal intensity:
- Low level of protein transferred from the gel to the membrane. This is particularly true for high molecular weight proteins. Reversible Protein Stain Kits for Nitrocellulose and PVDF membranes (Cat. Nos. 24580 and 24585, respectively) may be used to stain the bands on the membrane to indicate transfer efficiency.
- Over-diluted primary and secondary antibody probes. We suggest using the recommended dilution for the antibody probes for each substrate. More sensitive substrates will require a more dilute antibody solution.
- Short substrate incubation time. For best results, incubate the membrane for 5-15 min in the SuperSignal substrate.
I used the MemCode Reversible Protein Stain Kit but the stain is not completely reversible. Why is this?

Here are possible causes and solutions:

- Membrane was allowed to dry before reversing. Keep the membrane wet.
- High protein concentration in the sample. Extend incubation in the Stain Eraser up to 5 minutes. Reduce the protein concentration in the sample.
I used the MemCode Reversible Protein Stain Kit and my bands are faint or not visible at all. What is going on?

This is likely due to low amounts or no protein present in the sample. Please determine the protein concentration in the original sample.
What is the sensitivity of the MemCode Reversible Protein Stain Kit for Nitrocellulose Membranes?

The MemCode Reversible Protein Stain Kit for Nitrocellulose Membranes detects as low as 25 to 50 ng protein per band.
How should I store the Thermo Scientific MemCode Reversible Protein Stain Kit for Nitrocellulose Membranes and what is its shelf life?

We recommend storing the MemCode Reversible Protein Stain Kit for Nitrocellulose Membranes at room temperature where it is stable for a year.

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