K-Ras4B Mutant Protein: G12D+D38A (human recombinant with 6xHis-tag)

BlastR™ Lysis/Dilution Buffer Kit (50 lyses)
$0.00

The BlastR™ Buffer system allow efficient extraction of proteins from all cellular compartments (Fig. 1). BlastR™ lysis buffer system is compatible with standard colorimetric protein assays unlike Laemmli and 1% denaturing lysis buffer (Fig. 3) It should be noted that BlastR™ Buffer will efficiently break up protein:protein interactions and is therefore not suitable for co-IP applications.

NOTE: Lysates may become viscous due to the release of genomic DNA. The Lysate viscosity can be eliminated after passage over a BlastR™ filter (not included in this kit), providing a user friendly lysate for downstream applications including SDS-PAGE, western blotting, immunoprecipiatations (IPs). The BlastR™ filters are sold separately and can be used with any nuclear lysis buffer system to reduce lysate viscosity by removing genomic DNA from the sample (Fig. 2)

Product Uses Include 

  • Remove genomic DNA
  • Suitable for SDS-PAGE Application
  • Suitable for IP Applications

Sample Kit Components (50 lyses)

  • BlastR™ Lysis buffer 
  • BlastR™ Dilution buffer
 

 

Figure 1. Comparison of BlastR lysis buffer to alternative lysis
buffers. A431 cells were lysed with BlastR, RIPA, mPER, IP lysis, Denaturing(1% SDS), and Laemmli lysis buffers. All denaturing lysates had genomic DNA removed using BlastR filter. Isolation of proteins from the membrane,cytoplasmic, mitochondrial, and nuclear markers were determined using antibodies against the respective compartment marker proteins.

BLR03 Fig 1

Figure 2. BlastR lysis filter is effective at removing genomic DNA.  (A) A431 cells were lysed with a denaturing lysis buffer. Genomic DNA was removed or sheared with BlastR filter, syringe needle or soni-cation for 5, 10, 20, and 30 seconds. 2% of lysate was analyzed by  ethidi-um bromide, agarose gel electrophoresis. (B) Lysate from A431 cells lysed with a denaturing buffer was either unfiltered or filter with the BlastR filter.  Sample were separated with SDS-PAGE and visualized using Coomassie stain. (C) Duplicate samples from B were separated by SDS-PAGE, trans-ferred to PVDE, and EGFR protein was examined using an EGFR anti-

BLR03 Fig 2

Figure 3. BlastR lysis buffer characteristics. (A) A431 cells were lysed with BlastR, RIPA, mPER, IP lysis, Denaturing (1% SDS), and Laemmli lysis buffers. All denaturing lysates had genomic DNA removed using the BlastR filter. Coomassie stain was performed to obtain a protein isolation profile with these buffers. (B) Protein quantitation of RIPA, BlastR, 1% SDS, and Laemmli was performed using a standard colorimetric assay (ADV02). A titration of 5, 10, and 20 µL was performed to determine the 

BLR03 Fig 3

 

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