We obtained glycine, Tris, TritonX-100, DA, homovanillic acid (HVA), 3, 4-dihydroxyphenylacetic acid (DOPAC), MPTP, dithiothreitol (DTT), sodium dodecyl sulfate (SDS), and LY294002 from Sigma-Aldrich (St. Louis, MO, United States). From our institutional pharmacy, this study acquired Madopar (Shanghai Roche Led., Shanghai, China). Abcam (Dako, Cambridgeshire, United Kingdom) offered Rat anti- glyceraldehyde 3-phosphate dehydrogenase (GAPDH), anti-Akt (total) antibody, rabbit anti-BAD, rabbit anti-cleaved-Caspase-3 antibody, rabbit anti-phospho-Akt (Ser473) antibody, p-BAD, mTOR, p-mTOR, BDNF, CREB, and p-CREB antibodies, horseradish peroxidase (HRP)-conjugated anti-rabbit, and HRP-conjugated rat antibodies. All other chemicals exhibited great-purity analysis level and originated in Shanghai Chemical Reagent Co., Ltd. (Shanghai, China).
Rabbit anti bad
Manufactured by Abcam
Sourced in United Kingdom, United States
Rabbit anti-BAD is a primary antibody that recognizes the BAD (Bcl-2-associated death promoter) protein. BAD is a pro-apoptotic member of the Bcl-2 family of proteins and plays a role in regulating cell survival and apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions)
P bad, by Abcam (1 mentions)
Rabbit anti cleaved caspase 3 antibody, by Abcam (1 mentions)
Ly294002, by Merck Group (1 mentions)
P mtor, by Abcam (1 mentions)
Rabbit anti bax, by Abcam (1 mentions)
Image lab 5, by Bio-Rad (1 mentions)
Rabbit anti p p65 ser536, by Cell Signaling Technology (1 mentions)
Rabbit anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Mouse anti β actin, by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
Rabbit anti lc3b, by Abcam (1 mentions)
Rabbit anti pten, by Abcam (1 mentions)
Rabbit anti gapdh, by Abcam (1 mentions)
Ecl reagent, by Merck Group (1 mentions)
3 protocols using rabbit anti bad
1
Investigating Neurochemical Signaling Pathways
2
Protein Extraction and Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins from renal tissues were extracted using the RIPA lysis buffer (Thermo Fisher, USA) and quantified using the BCA Protein Assay Kit (Thermo Scientific, USA). Equal concentrations of proteins were separated on 10 or 15% SDS-polyacrylamide gels and were subsequently transferred to a PVDF membrane (Millipore, USA). The membranes were blocked using 5% nonfat milk for 60 min, under room temperature, then incubated overnight with primary antibodies at 4°C. In order to bind primary antibodies, HRP-conjugated secondary antibodies were incubated for 60 min under room temperature. Blots were detected and visualized with Clarity Western ECL Substrate (Bio‐Rad) under a MP imager (Bio‐Rad developed). Density analysis of the protein bands was performed using Image Lab 5.1 which was used for (Bio-Rad, USA). The primary antibodies used in the study were against the proteins and included the following: rabbit anti-cleaved caspase-3 (1 : 250) (Cell Signaling Technology, USA), rabbit Anti-Bad (1 : 1000), rabbit Anti-Bax (1 : 1000) (Abcam, USA), rabbit Anti-GAPDH (1 : 1000) (Cell Signaling Technology, USA), rabbit Anti-p-p65(Ser536) (1 : 1000) (Cell Signaling Technology, USA), and mouse anti-β-actin (1 : 5000) (Sigma-Aldrich, USA).
3
Protein Expression Analysis in MPC-5 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The MPC-5 cells in each group were rinsed twice with PBS, and the PBS was aspirated prior to the addition of RIPA cell lysis solution (Thermo Fisher Scientific, Inc.). The cells were then shaken on ice for 30 min and lysed by pipetting. Total protein concentration was measured using the BCA method and the proteins were denatured at 100˚C for 10 min. Total proteins (30 µg) were separated by SDS-PAGE on 12% gels and were transferred to PVDF membranes. Blocking was performed with 5% nonfat milk powder at 25˚C on a shaker for 2 h, after which, dropwise rabbit anti-DJ-1 (1:1,000; cat. no. ab76008; Abcam), rabbit anti-PTEN (1:1,000; cat. no. ab137337; Abcam), rabbit anti-LC3B (1:1,000; cat. no. ab192890; Abcam), rabbit anti-Bad (1:1,000; cat. no. ab32445; Abcam) or rabbit anti-GAPDH (1:1,000; cat. no. ab9485; Abcam) antibodies were added, and incubated overnight at 4˚C on a shaker. The horseradish peroxidase-conjugated secondary antibody (1:1,000; cat. no. ab6721; Abcam) was then added for 1 h at room temperature on a shaker. Subsequently, 1 ml western blot ECL reagent (MilliporeSigma) was added to the blots, which were exposed, and images were captured. The exposed image was scanned using ImageJ (V1.8.0; National Institutes of Health) and the gray value of the protein band was analyzed.
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!