Samples were lysed in RIPA buffer (P0013B, Beyotime) containing protease inhibitor cocktail (P8340, Sigma) and PMSF (A610425-0005, Sangon Biotech). Proteins were separated by SDS–PAGE and transferred onto polyvinylidene difluoride membranes. Blocking and antibody incubations were performed in 5% BSA or non-fat dry milk. Anti- PPARγ (16,643-1-AP, 1:1000), anti-FABP4 (12,802-1-AP, 1:1000), anti-PERILIPIN2 (15,294-1-AP), anti-PGC1α (66,369-1-Ig) and anti-UCP1 (23,673-1-AP) were purchased from Proteintech. Anti-β-actin (A5441, 1:1500) was purchased from Sigma-Aldrich. Antibody detection reactions were developed by enhanced chemiluminescence reagent (Millipore) and imaged using the MiniChemi 610 imaging system (Sage Creation). Quantification was done using Lane 1D software (Sage Creation).
Minichemi 610 imaging system
Manufactured by Sagecreation
Sourced in Germany, China
The MiniChemi 610 is a compact imaging system designed for the capture and analysis of chemiluminescent and fluorescent signals. It features a high-resolution CCD camera, adjustable lighting, and dedicated software for image acquisition and processing.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence reagent, by Merck Group (1 mentions)
Lane 1d software, by Sagecreation (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Sangon (1 mentions)
Anti ucp1 23673 1 ap, by Proteintech (1 mentions)
Anti fabp4, by Proteintech (1 mentions)
Anti pparγ 16643 1 ap, by Proteintech (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Mmp9 antibody, by Abcam (1 mentions)
β actin antibody, by Abcam (1 mentions)
Protease phosphatase inhibitor, by Beyotime (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Applygen (1 mentions)
3 protocols using minichemi 610 imaging system
1
Western Blot Analysis of Adipocyte Markers
2
MMP-9 Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression of MMP-9 protein was detected by Western Blot. Cells were lysed in lysis buffer supplemented with a protease inhibitor cocktail. Protein lysates mixed with loading buffer were heated at 95 °C for 5 min and were then loaded onto 10 % SDS- PAGE gels. The proteins were separated out by electrophoresis and transferred to a polyvinylidene difluoride (PVDF) membrane. Membranes were blocked with 5 % BSA for 1 h at room temperature, incubated with MMP-9 antibody (1:1000, Abcam, Massachusetts, USA) and β-actin antibody (1:5000, Abcam) overnight at 4 °C, then incubated with HRP-conjugated secondary antibody for 1 h at room temperature. Blots were detected by the enhanced chemiluminescence (ECL) system (Millipore, Germany) and imaged with the Mini Chemi610 Imaging System (Sagecreation, Beijing, China).
3
Evaluating PD1 Gene Recombinant Exosomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For identifying PD1 gene recombination engineered exosomes, Western blot assay was performed to evaluate the PD protein expression. Briefly, Exo, PD1 Exo, PD1-Imi Exo or gene recombinant HEK 293T cells were collected, and lysed in RIPA buffer (Applygen, Beijing, China) with protease phosphatase inhibitor (Beyotime, Shanghai, China) at 4 °C for 1 h. The lysed mixture was then centrifuged at 12000 rpm at 4 °C for 20 min for collecting its supernatant. Afterwards, the protein concentration in the supernatant was quantified by BCA assay kit, and further separated by 4%–20 % precast sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel. The separated proteins were transferred onto polyvinylidene fluoride (PVDF) membranes (Merck-Millipore, Darmstadt, Germany) and blocked with 5 % bovine serum albumin (BSA) in TBS-T (Tris buffered saline with Tween 20) solution. After incubating with the appropriate primary antibodies at 4 °C overnight, the PVDF membranes was further incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies at 25 °C for 1 h. Finally, the blots were observed by a MiniChemi610 imaging system (Sage Creation, Beijing, China).
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!