Uniblue A sodium 

Uniblue A sodium is a reactive protein stain that can be used in the covalent pre-gel staining of the protein (Ex=594 nM)^[1].

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1].
1. Prepare the derivatization buffer: 100 mM NaHCO₃, 10% SDS, pH 8-9.
2. Add 10 μL of 200 mM Uniblue A solution (dissolved in derivatization buffer) to 90 μL protein solution.
3. The samples are heated at 100°C for 1 min to perform the staining.
4. Add 100 μL of reducing solution composed of 10% glycerol and 20 mM dithiotreitol (DTT) in 200 mM Tris buffer with pH 6.8.
5. Excess Uniblue A reacts with Tris, resulting in a blue compound, serving as running front indicator in electrophoresis.
6. The sample is heated another minute at 100°C in order to achieve efficient reduction and allowed to cool to room temperature.
7. Add 20 μL alkylation solution containing 550 mM iodoacetamide (IAA), incubated for 5 minutes.
8. the samples are subjected to SDS-PAGE.
Note: Dry protein samples or samples in compatible buffers (i.e. free of amines) can be diluted directly with the derivatization buffer solution to 5 mg/mL. In other cases, a prior trichloroacetic acid (TCA)/acetone precipitation or buffer exchange by ultrafiltration is recommended (see below).
As positive control, bovine serum albumin (BSA) in derivatization buffer is used at a concentration of 10 mg/mL.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

506.48

C₂₂H₁₅N₂NaO₇S₂

14541-90-3

594

555

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Marco A Mata-Gómez, et al. Accelerated identification of proteins by mass spectrometry by employing covalent pre-gel staining with Uniblue A. PLoS One. 2012;7(2):e31438.  [Content Brief]