What gel to use:

Product Name

Manual Electrophoresis

Protogel        37.5:1 30%

1 D and 2 D gels,            Iso-Electric Focusing, Gradient gels etc

Accugel            29:1 40%

As above, also used for smaller proteins

Acrylagel        30%

As above

Bis-Acrylagel  2%

As above

ProtoPrep

Sample recovery 10-300kD range
ProtoGel
Order No. EC-890

Price/1Liter  :     -15 %   = 73 Euro

ProtoGel™

37.5:1 Acrylamide to Bis-Acrylamide
Stabilized Solution

National Diagnostics' ProtoGel is a stabilized ready-to-use, 30% (w/v) acrylamide solution, with the cross-linker, methylene bisacrylamide (37.5:1 ratio), in distilled/deionized water. ProtoGel results in the same polyacrylamide gels that you are now accustomed to preparing, because it is identical to your own solutions. With ProtoGel, you can now prepare the same gel faster, easier, safer, and more reliably. All you need to do is pour and use.

ProtoGel is supplied in 450 ml and 1 Liter bottles containing 300 grams of acrylamide and 8 grams of methylene bisacrylamide per liter of solution.

Storage: ProtoGel is stable for 24 months when stored tightly capped, in a dark area at room temperature (20°C).
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ProtoGel Buffer
Order No. EC-892
450 ml
1 Liter
ProtoGel Stacking Buffer
Order No. EC-893
200 ml

ProtoGel Buffer Solutions

ProtoGel Buffer (sieving gel buffer) and ProtoGel Stacking Buffer compliment National Diagnostics' ProtoGel by providing ready-to-use solutions of non-monomeric components. By using ProtoGel Buffer and ProtoGel Stacking Buffer, all you need to add is ammonium persulfate and TEMED to complete the preparation of SDS-PAGE separating and stacking gels. ProtoGel Buffer and ProtoGel Stacking Buffer are designed as optional replacements for the additional solid components usually required when preparing ProtoGel. ProtoGel Buffer can be used to prepare any percentage gel desired.

ProtoGel Buffer is supplied in 450 ml and 1 Liter bottles forming gels of .375M Tris-HCL and 0.1% SDS, pH 8.8. ProtoGel Stacking Buffer is supplied in 200 ml bottles, forming gels of .125M Tris and 0.1% SDS, pH 6.8

Storage: ProtoGel Buffer and ProtoGel Stacking Buffer are stable for 12 months when stored tightly capped, in a dark area at room temperature (20°C).

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Staining with InsiteTM Kit
  • Staining Occurs During the Run
  • Requires No Denaturing Fixatives
  • Mild Conditions Optimize Protein Recovery
  • 10ng Sensitivity in 30 Minutes
 
Applications


Post-electrophoretic protein detection

Denaturing electrophoresis of proteins

Western blotting

Insite is a run-time protein visualization system incorporating a unique dye/buffer combination which stains proteins during the gel run. The Insite System has been optimized for staining standard minigels run using the Laemmli buffer system, such as with National Diagnostics' ProtoGel, ProtoGel Resolving Buffer, and ProtoGel Stacking Buffer. Insite will detect as little as 10ng of protein per band.

Insite also functions optimally in combination with the ProtoPrep II Kit, National Diagnostics' meltable acrylamide matrix for the electrophoretic separation and purification of proteins.

The Insite protocol is fast, easy, and mild:

  1. Prepare 150 ml of Insite Detection Reagent by diluting 15 ml of 10X Insite Buffer Concentrate to 149 ml with deionized water and adding 1 ml of Insite Dye Concentrate.
  2. Mount the gel in the apparatus.
  3. Fill the lower buffer chamber with 1X Tris Glycine SDS (EC-870) (0.192M Glycine, 0.025M Tris, and 0.1% SDS).
  4. Fill the upper buffer chamber with the Insite Detection Reagent.
  5. Run the gel under standard voltage and temperature conditions. Typical running conditions are 175 volts for 1 hour.
  6. Remove the gel from the apparatus. [The gel may be observed immediately using a UV transilluminator (see below) for the detection of bands containing more than 300ng of protein.]
  7. Destain the gel in 50 ml deionized water for 15 minutes. Repeat the wash in fresh deionized water for another 15 minutes.
  8. Observe the gel on a transilluminator under ultraviolet illumination (302nm). Protein bands fluoresce yellow-orange. If excessive background is observed, wash the gel in an additional deionized water bath for 5-10 minutes.
  9. Bands may be photographed using the #8 yellow filter used for coomassie stains. Note that background will increase as the gel is allowed to stand and the surface dehydrates. This background is easily removed by a brief wash in deionized water.
 
Insite Kit
Order No. EC-759
1 Kit Stains 30 MiniGels

+ Free samples of Western Blot Recycling Kit Cat #90100

Do NOT discard precious blots!! Strip Antibodies and Re-use Protein blots In Just 15-30 min.!!

Western blotting is a commonly used technique to study protein structure and function. Typically, protein samples are electrophoresed on SDS-gels and transferred to solid support (nitrocellulose or PVDF membranes) for subsequent probing with specific antibodies. Unlike advances made in RNA/DNA blotting, it has been difficult to strip antigen/antibodies and reuse blots.

Recycling of blots offers many advantages:

The idea of reusing protein blots derives strength from the fact that antigen-antibody complexes (e.g., immunoaffinity columns) can be disrupted and the immunoaffinity column reused many times. However, commonly employed elution conditions in immunoaffinity techniques (low or high pH, use of chaotropic agents) have not been effective in stripping antibodies from protein blots.

ADI has now developed specially formulated solutions that effectively and almost quantitatively strip antibodies under gentle conditions. It offers the following advantages:

Advantages of New Antibody Strip Solution

1. Strip antibodies at room temperature (no heating is required).
2. No pungent smelling, mercaptoethanol.
3. Strip solution comes in 10X solution and it is ready to use. Just soak blots in the soln for 10-15 min.
4. Blots are reusable within 15-60 min after a short incubation in blocking buffer (provided in the kit).

Ordering information - Click Here For On line Ordering

Western Blot Recycling Kit (reagents for stripping 20-30 std. or 50 mini blots) Cat # 90100; 250 Euro / 205 $

Information Request Form.

Send -35 % promotion (until 15/01/2003) + free Western Blot Recycling kit
Send Insite Staining information
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