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Can I fix my gels in ethanol/acetic acid rather than methanol/acetic acid prior to staining with SYPRO Ruby Gel Stain? Product FAQ

Answer

Yes, but ethanol/acetic acid will produce ethyl acetate, which has a strong odor, so you should do this fixation in a fume hood.

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

Answer Id: E11134

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Do you recommend performing an alcohol/acetic acid fixing step prior to staining with SimplyBlue SafeStain? Product FAQ

Answer

SimplyBlue SafeStain does not contain acetic acid or methanol. An alcohol/acetic acid fixing step prior to staining with SimplyBlue SafeStain is not required or recommended.

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

Answer Id: E11065

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What solvent is Collagen I, bovine (Cat. No. A1064401) shipped in? Product FAQ

Answer

Collagen I, bovine (Cat. No. A1064401) is shipped in 20 mM acetic acid.

Answer Id: E21070

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What solvent can be used to dissolve Blasticidin? Product FAQ

Answer

Blasticidin can be dissolved in water or acetic acid. Make stocks of 5-10 mg/ml.

Answer Id: E3690

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When staining IEF gels, can a normal acetic acid fix be used instead of sulfosalicylic acid? Product FAQ

Answer

No. An acetic acid fix is not strong enough. However, a 12% trichloroacetic acid (TCA) fix can be substituted.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Answer Id: E10651

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What is the 1X buffer composition of your TAE electrophoresis buffer (Cat. No. B49)? Product FAQ

Answer

The 1X buffer composition is as follows: 40 mM Tris, 20 mM acetic acid, 1mM EDTA.

Answer Id: E13215

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Why is SimplyBlue SafeStain a "safe" stain? Product FAQ

Answer

SimplyBlue Safestain is specially designed for safe, non-hazardous disposal. It does not contain methanol or acetic acid and does not require the use of methanol or acetic acid which must be disposed of as hazardous waste.

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

Answer Id: E11066

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Acetic Acid in Canned Products Product Literature

Is collagen I provided lyophilized or as a liquid? What is the buffer composition if it is a liquid? Product FAQ

Answer

Collagen I is provided as a liquid, suspended in 20 mM acetic acid solution. If difficulties are encountered with the preparation, the collagen can be diluted in 1:2, first in 20 mM acetic acid, pH 3.5 before further diluting according to the protocol being used.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E11807

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How do I prepare a solution of Coomassie R-250 dye? Product FAQ

Answer

Add 100 mL of glacial acetic acid to 450 mL distilled water.
Dissolve 3 g of Coomassie R-250 dye in 450 mL methanol.
Mix the acetic acid and methanol solutions.
Filter the solution before use.

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

Answer Id: E13303

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What do you recommend for transferring IEF gels? Product FAQ

Answer

For IEF gels, we recommend using an acetic acid transfer buffer. The IEF gels are 5% polyacrylamide and transferring them in a basic buffer leads to substantial hydrolysis and damage to the gel. The following protocol prevents the gel from hydrolyzing and is especially effective for basic proteins because of the low pH of the transfer buffer:

After the run, equilibrate the gel in 0.7% acetic acid (0.7% acetic acid in water, pH 3.0) for 10 minutes. Chill the 0.7% acetic acid that will be used as the transfer buffer. Assemble the gel/membrane sandwich in reverse order so that the membrane is in contact with the side of the gel facing towards the cathode (-). This is opposite from the typical western blot with SDS, where the negatively charged protein will migrate toward the anode (+) side during the transfer. Transfer for one hour at 10 volts.

Tip for handling the IEF gel: The 5% polyacrylamide IEF gels tend to be sticky. While the gel is floating in the equilibration solution, submerge the filter paper underneath the gel. When the gel is in the correct position, lift up on the filter paper so the gel attaches to it. Floating the gel over the filter paper avoids the need to handle the gel and prevents the gel from getting stuck onto the filter paper before it is in its proper position.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Answer Id: E10652

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How do you Western transfer/blot vertical IEF gels? Product FAQ

Answer

For IEF gels, we recommend using an acetic acid transfer buffer. The IEF gels are 5% polyacrylamide and transferring them in a basic buffer leads to substantial hydrolysis and damage to the gel. The following protocol prevents the gel from hydrolyzing and is especially effective for basic proteins because of the low pH of the transfer buffer:

1) After the run, equilibrate the gel in 0.7% acetic acid (0.7% acetic acid in water, pH 3.0) for 10 min.

2) Chill the 0.7% acetic acid that will be used as the transfer buffer.

3) Assemble the gel/membrane sandwich in reverse order so that the membrane is in contact with the side of the gel facing towards the cathode (-). This is OPPOSITE from the typical western blot with SDS-PAGE, where the negatively-charged protein will migrate toward the anode (+) side during the transfer.

4) Transfer for one hour at 10 V.

Tip for handling the IEF gel: the 5% polyacrylamide IEF gels tend to be sticky. While the gel is floating in the equilibration solution, submerge the filter paper underneath the gel. When the gel is in the correct position, lift up on the filter paper so the gel attaches to it. Floating the gel over the filter paper avoids the need to handle the gel and prevents the gel from getting stuck onto the filter paper before it is in its proper position.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Answer Id: E3560

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Do I have to fix my gel in 50% methanol/7% acetic acid, or can other fixative solutions be used prior to staining with SYPRO Ruby Gel Stain? Product FAQ

Answer

No, other commonly used gel fixatives can be used, including reducing the methanol concentration to 10% methanol in 7% acetic acid. SYPRO Ruby stain itself will fix proteins in the gel and there is no need for a separate fixation step to stain proteins with reasonably good results. The 50% methanol/7% acetic acid fixation recommended in the protocol has been determined to best remove residual SDS from the gel matrix and thus give the lowest background and optimal sensitivity compared to other fixation methods. Reduced methanol concentrations could result in a heavily stained SDS front at the bottom of the gel, which will reduce detection sensitivity for low molecular weight proteins running near this region.

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

Answer Id: E11135

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I would like to use Collagen I Rat Protein, Tail (Cat. No. A1048301) for coating cell culture dishes. In the first step of the thin coating protocol, what should I use to dilute the 17.4 M acetic acid stock solution to 20 mM? Product FAQ

Answer

You can just use water to dilute the acetic acid stock solution to 20 mM.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17377

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What is the storage buffer composition for the resin slurry in the columns within the High-Select Fe-NTA Phosphopeptide Enrichment Kit (Cat. No. A32992)? Product FAQ

Answer

The resin slurry is in 5% acetic acid.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Answer Id: E19607

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