Gingival samples were treated with RIPA buffer (pH 7.0) (R0278-50ML, Sigma-Aldrich, Inc. USA) containing Halt™ Protease and Phosphatase Inhibitor Cocktail, EDTA-free (1:100, 78,441, Thermo Scientific™, USA). Prestained Protein Standards, Precision Plus Protein™ Kaleidoscope™ (1,610,375, Bio-Rad Laboratories, Inc., USA) were used as molecular weight markers. The samples were electrophoresed on SDS-PAGE (8% separating and 4% stacking gels) and then electrophoretically transferred to nitrocellulose membranes. Western blot analysis was carried out as described by us previously.36 (link) Specific immunoreactivity was visualized as dark bands against a clear background, and the membranes were scanned with Invitrogen™ iBright™ FL1000 Western Blot Imaging Systems (Thermo Fisher Scientific, Inc., USA) for imaging and documenting chemiluminescent Western blot bands. The densitometric units were measured in the linear range of immunoreactivity for TLR-2 and p38 MAPK expressions by Image J analysis software.
R0278 50ml
Manufactured by Merck Group
Sourced in United States
R0278-50ML is a laboratory reagent product from Merck Group. It is a 50 ml solution for scientific research and analysis purposes. The core function of this product is to serve as a reagent for various laboratory procedures. No further details on the intended use or composition of this product can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Precision plus protein kaleidoscope, by Bio-Rad (1 mentions)
Invitrogen ibright fl1000 western blot imaging systems, by Thermo Fisher Scientific (1 mentions)
Halt protease and phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Anti gapdh kc5g4, by Kangchen Biotech (1 mentions)
Ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Hybond ecl nitrocellulose membrane, by Cytiva (1 mentions)
Bradford reagent, by Merck Group (1 mentions)
Bca assay, by Thermo Fisher Scientific (1 mentions)
P8340 5ml, by Merck Group (1 mentions)
Ipvh00010, by Merck Group (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Dc protein assay, by Bio-Rad (1 mentions)
4 protocols using r0278 50ml
1
Gingival Tissue Protein Analysis
2
Quantitative Protein Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein isolation was performed through cell lysis buffer (R0278-50ML, Sigma-Aldrich Co., USA) with their concentrations quantified by Bradford reagent (B6916-500ML, Sigma-Aldrich Co., USA). After boiling, 30 μg of proteins were separated by polyacrylamide gel electrophoresis and transferred to a Hybond ECL nitrocellulose membrane (Amersham Biosciences, Arlington Heights, IL, USA). Membranes were then blocked with 5% low-fat dried milk in phosphate buffered saline containing 0.1% Tween-20 and subsequently incubated for 1 h at room temperature with 1:1,000 dilutions of the primary ZNF703 antibody. Mouse monoclonal anti-GAPDH (KC5G4, KangChen Biotech, Shanghai, China) was used as a loading control. Membranes were then exposed to ECL western blotting substrate (32209, Thermo Fisher Scientific Inc. Waltham, USA) after incubating for 1 h with 1:1,000 dilution of the second antibody in darkness.
3
Western Blot Analysis of T1 and T2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T1 and T2 cells were lysed and protein was isolated using Radioimmunoprecipitation assay (RIPA) lysis buffer (R0278-50ML, Sigma-Aldrich) with protease inhibitor (P8340-5ML, Sigma-Aldrich). The protein concentrations were quantified using BCA assay (23225, Thermo Fisher Scientific). 25 µg of total protein lysate was loaded on to 10% SDS gel and western blotting was performed as previously described (42 (link)). Antibodies used in the study are listed in Supplementary Table 5
4
Mertk Protein Extraction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cell lysates, cells were lysed with a RIPA buffer (R0278-50ML, Sigma Aldrich, Burlington, MA, USA) and a Halt 100× protease/phosphatase inhibitor cocktail (1861281, Thermo Scientific, Rockford, IL, USA). After centrifugation, a final protein concentration of 2–8 mg/mL was calculated using the DC Protein Assay (500-0116, Bio-Rad, Hercules, CA, USA). For the collection and analysis of soluble Mertk in the cell culture media, media were concentrated using a 10 kDa molecular weight cut-off centrifugal filter (UFC501096, Amicon, Darmstadt, Germany). For both the cell lysate and concentrated media, a 6 X SDS protein-loading buffer was prepared (341 mM of Tris-HCl, 8.2% SDS, 45.5% glycerol, 0.03% bromophenol blue, 9% β-mercaptoethanol) and added to the prepared protein. Prepared samples were boiled for 5 min, run via SDS-PAGE electrophoresis, and transferred to the PVDF membrane (IPVH00010, Millipore, Cork, Ireland) for analysis.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!