Effects of the modification of transfer buffer composition and the renaturation of proteins in gels on the recognition of proteins on Western blots by monoclonal antibodies.monoclonal antibodiesimmunochemical methodsWestern blotsblotting to nitrocellulose...
Western blot analysis Total proteins were extracted from HCC tissues or cell lines using RIPA buffer (Beyotime, Shanghai, China) with a complete protease inhibitor cocktail and phosphatase inhibitors (Roche, Basel, Switzerland). Aliquots of protein extracts were separated on 12% SDS-PAGE and transf...
Cells were washed and resuspended in lysis buffer C. The resuspended cells were lysed by stirring for 3 to 4 h in the presence of DNase I (20 μg/ml) before clearance at 15,000 × g for 30 minutes. Cleared lysate was filtered and passed over GSTrap or MBPTrap column (Cytiva) ...
PfSTART1 protein was dialysed extensively against 50 mM NaPO4-3 (pH 7.4), 150 mM NaCl, before diluting to 90 μM in the same buffer with 2% (v/v) DMSO (syringe sample). The first injection was 0.4 μL over a 0.8 s duration, and the remaining 19 injections were 2...
The cells were collected 2 days after transfection, washed twice with PBS and lysed with cell lysis buffer (Cell Signaling Technology, Danvers, MA, USA). Western Blot was used to verify the expression of the ZIKV-prME protein from the harvested cell lysate and the immune specificity of the ...
The compositions can include the lipid-based carrier, material to be introduced into a cell, and BDM in a carrier, such as a cell compatible buffer. The composition can be diluted for application to cells, or can be provided at “ready to use” concentrations for application to cells....
For this, EVs were dissolved in SDS-sample buffer, and the particle concentration was adjusted based on the protein concentration measured by the BCA method using bovine serum albumin (BSA) as a standard. For preparation of cell lysates of P29 and A11 cells, they were lysed in RIPA buffer ...
To form these liposomes, one mixes the DNA of an expression vector which expresses the gene to be transferred with lipid, such as N-[1-(2,3,-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) in a suitable buffer, such as Hepes buffered saline. This causes the spontaneous ...
The sample was fixed with 2% glutaraldehyde and 2% paraformaldehyde in 0.1 M sodium cacodylate buffer at pH 7.3. After two washes in distilled H2O, the samples were stained with 2% methylamine tungstate for 45 s and air-dried. Finally the samples were observed in a Zeiss scanning electron ...
Western blotting Cells were seeded on aligned scaffolds for 2 days, dynamically loaded for 15 min at 1 Hz to 3% strain, washed once with PBS, and lysed in RIPA buffer (50 mM Tris-HCl pH 8.0, 150 mM sodium chloride, 1.0% Triton X-100, 0.1% SDS, with 1% protease inhibitor cocktail ...