Bioactivity | 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]. |
Invitro | 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. |
Name | Uniblue A sodium |
CAS | 14541-90-3 |
Formula | C22H15N2NaO7S2 |
Molar Mass | 506.48 |
Transport | Room temperature in continental US; may vary elsewhere. |
Storage | Please store the product under the recommended conditions in the Certificate of Analysis. |
Reference | [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. |