Tissues and cells were lysed with RIPA buffer containing protease inhibitor (Bimake, #B14002) and phosphatase inhibitor (Bimake, #B15003). Bicinchoninic acid (BCA) protein assay kit (Yeasen, #20201ES90) was used to detect protein concentration. Equal quantity of proteins was separated by SDS‐PAGE and transferred to PVDF membrane. The membrane was blocked with 5% bovine serum albumin (Yeasen, #36101ES80) for 1–3 h at room temperature, and subsequently incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies for 1–3 h at room temperature. Next, the electrochemiluminescence reagents (Yeasen, #36208ES76) were used to detect the antibody signals, and quantitative analysis of the developed results was performed by ImageJ software using vinculin as an internal reference to calculate the relative expression levels of target protein. Primary antibodies of IMPA1(Abcam, #ab184165), Vinculin (Sigma, #V9131), and HA (CST, #3724S) were used in this study. Other antibody information is shown in Table S5 .
Bicinchoninic acid bca protein assay kit
Manufactured by Yeasen
Sourced in China
The Bicinchoninic acid (BCA) protein assay kit is a colorimetric detection and quantification method used to measure the total protein concentration in a sample. The kit utilizes the reaction between protein and an alkaline copper tartrate solution, which results in the reduction of Cu2+ to Cu+. The reduced copper then interacts with bicinchoninic acid, producing a purple-colored complex that can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Yeasen (1 mentions)
Electrochemiluminescence reagents, by Yeasen (1 mentions)
Oligomycin, by Merck Group (1 mentions)
Antimycin a, by Abcam (1 mentions)
Mitotracker green fm, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 annexin 5 dead cell apoptosis kit, by Thermo Fisher Scientific (1 mentions)
Fluoro carbonyl cyanide phenylhydrazone (fccp), by Merck Group (1 mentions)
Rotenone, by Merck Group (1 mentions)
Dcfh da, by Yeasen (1 mentions)
2 protocols using bicinchoninic acid bca protein assay kit
1
Western Blot Analysis of Protein Expression
2
Comprehensive Metabolic Profiling of Cellular Stress
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GAS with a purity >98% was purchased from Nature Standard Company (Shanghai, China). SRB (sulforhodamine B), TCA (trichloroacetic acid), oligomycin, FCCP (2- [2-[4-(trifluoromethoxy)phenyl]hydrazinylidene]-propanedinitrile), and rotenone were purchased from Sigma-Aldrich (St. Louis, MO, USA). MitoTracker Green FM and the Alexa Fluor 488 Annexin V/Dead Cell Apoptosis Kit were purchased from Thermo Fisher Scientific (Waltham, MA, USA). DCFH-DA (2′,7′-dichloro-fluorescin diacetate) and the bicinchoninic acid (BCA) protein assay kit were obtained from Yeasen (Shanghai, China). Antimycin A was purchased from Abcam Biotechnology (Cambridge, MA, USA). The reverse-transcription kit was obtained from TaKaRa (Shiga, Japan). The FastStart Essential DNA Green Master for PCR was purchased from Roche (Basel, Switzerland). Antibodies, except where specifically noted, were purchased from Cell Signaling Technology (Danvers, MA, USA).
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!