Streptococcus pyogenes M1 CRISPR-Cas9 Baculovirus-Insect Overexpression Lysate: Product Information
This Streptococcus pyogenes M1 CRISPR-Cas9 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of CRISPR-Cas9 protein (Cat: 40572-A08B) from the overexpression lysate was verified.
Streptococcus pyogenes M1
A DNA sequence encoding the s. pyogenes serotype M1 cas9 (WP_010922251.1) (Met1-Asp1368) was expressed with a nuclear localization sequence (NLS) at the N-terminus, two NLSs and a polyhistidine tag at the C-terminus.
The recombinant s. pyogenes serotype M1 cas9 consists of 1407 amino acids and predicts a molecular mass of 163 kDa.
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Successfully added to cart Please enter catalog numberSubmitted successfullyNetwork ErrorPlease enter your company namePlease enter your namePlease enter your emailPlease enter a valid email addressPlease enter some messageNot found.