Semba Biosciences offers the following convenient reagents for protein sample preparation.

SembaSonic™ 10X Protein Extraction Reagent, a mild detergent-based solution for efficient and gentle protein extraction without harsh mechanical, chemical, and/or enzymatic treatments. Lyse-Aid Recombinant Lysozyme facilitates bacterial cell lysis alone or in combination with SembaSonic 10X Protein Extraction Reagent. The popular Benzonase Nuclease is available for effective removal of nucleic acids from protein preparations. Sonicase™ Non-detergent Protein Extraction Reagent is formulated to address the need for efficient extraction of proteins for structural studies and for other detergent-prohibitive applications. SembaChrom Imidazole Solution is a specially-treated concentrate with low UV absorbance which facilitates detection of recombinant proteins purified using immobilized metal affinity chromatography (IMAC).

SembaSonic™ Protein Extraction Reagent

A mild, efficient, detergent-based protein extraction reagent for reliable preparation of soluble protein fractions

SembaSonic 10X Protein Extraction Reagent

SembaSonic 10X Protein Extraction Reagent is a proprietary concentrate containing a non-ionic detergent designed for preparation of cell lysates from E. coli. When used at a 1X final concentration, soluble proteins are gently released without denaturation or interference with activity. When combined with a buffer such as Tris or phosphate, Lyse-Aid™ Recombinant Lysozyme, and Benzonase® Nuclease, bacterial cell walls are effectively disrupted and nucleic acids are degraded to produce a non-viscous protein extract suitable for assays or chromatography. SembaSonic provides an easy, fast and economical substitute for physical disruption such as sonication or French Press homogenization. SembaSonic is ideal for rapid preparation of bacterial cell lysates for purification of recombinant proteins with Semba’s Octave™ Chromatography System. Use 5 ml 1X SembaSonic in a desired buffer per each gram of cell paste.

Sonicase™ Non-detergent Protein Extraction Reagent

Formulated for detergent-free, rapid, and efficient extraction of bacterial proteins

Sonicase Non-detergent Protein Extraction reagent is designed for preparation of cell lysates from frozen E. coli cell pellets. The reagent combines Lyse-Aid™ Recombinant Lysozyme and Benzonase® Nuclease in concentrations optimized for efficient cell lysis and nucleic acid degradation in the absence of any detergents, making it ideal for protein preparation for structural studies and other detergent-prohibitive applications. Sonicase provides an easy substitute for physical disruption such as sonication or French Press homogenization. Just resuspend the frozen bacterial cell pellet in a desired buffer (7 ml/g wet cell paste), add 20 µl Sonicase per gram cell paste, incubate 10-20 min, and centrifuge to remove debris.

Benzonase Nuclease

Dramatically reduce the viscosity of protein lysates, with a rapid, enzymatic treatment

Benzonase is ideal for a variety of applications such as purification of recombinant proteins from cell extracts where complete digestion of nucleic acids is desirable. This enzyme is a genetically engineered endonuclease from Serratia marcescens produced and purified from an E. coli strain, and is widely used to reduce the viscosity of cell extracts and remove interference from nucleic acids in downstream procedures. Benzonase attacks and degrades all forms of DNA and RNA (single stranded, double stranded, linear and circular) and is effective over a wide range of conditions. Upon complete digestion, all free nucleic acids present in solution are converted to 5’-monophosphate terminated oligonucleotides 2-5 bases in length. The enzyme is available in two grades, purity > 99% and purity > 90%. Both preparations possess exceptionally high specific activities and are supplied free from measurable protease activities. Both grades are available at a high concentration of 250 U/µl.

Unit definition: One unit will digest sonicated salmon sperm DNA to acid-soluble oligonucleotides equivalent to a ΔA260 of 1.0 in 30 min at pH 8.0 at 37 °C (reaction volume 2.625 ml).

Lyse-Aid Recombinant Lysozyme

A great tool for enzymatic lysis of the E. coli cell walls

Lyse-Aid™ Recombinant Lysozyme is a highly purified and stabilized lysozyme for lysis of gram-negative bacteria, such as E. coli. The enzyme catalyses the hydrolysis of N-acetylmuramide linkages in bacterial cell walls. The specific activity of Lyse-Aid Recombinant Lysozyme (1,700 KU/mg) for E. coli lysis is 250 times greater than of chicken egg white lysozyme. Lyse-Aid is optimally active at physiological pH and enhances the efficiency of protein extraction with SembaSonic™ Protein Extraction Reagent at very low concentration (3-5 KU/g cell paste). The product is supplied as a ready-to-use solution at a concentration of 30 KU/μl in 50% glycerol containing 50 mM Tris-Cl, 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% TRITON® X-100, pH 7.5.

Unit definition: One unit of Lyse-Aid Recombinant Lysozyme is defined as the amount of enzyme necessary to cause a decrease of 0.025 A450 unit per minute at 25ºC in a 1-ml suspension (1 mg/ml) of BL21 cells in 0.5X SembaSonic Reagent diluted with 50 mM Tris-Cl, pH 7.5.

Imidazole, SembaChrom™ Grade, 2.5 M Solution

The convenience and assurance of pure, low UV-absorbing, qualified & pH adjusted reagent for IMAC protein purification

Imidazole, SembaChrom™ Grade, is a special preparation of imidazole having low UV absorbance. Solutions containing imidazole are commonly used for the purification of histidine-tagged proteins by immobilized metal affinity chromatography (IMAC). Many commercial imidazole preparations contain UV-absorbing contaminants that interfere with spectrophotometric readings of protein solutions at 280 nm. SembaChrom Grade Imidazole has been processed to remove these contaminants, resulting in negligible background levels. The ready-to-use 2.5 M solution has been adjusted to pH 8.0 and filtered for convenient preparation of IMAC buffers.