Cell lysates were harvested in radioimmune precipitation assay buffer containing 150 mM NaCl, 10 mM EDTA, 50 mM Tris‐HCl at pH 7.4, 1% NP‐40, 0.004% sodium azide, 0.5% sodium deoxycholate, 0.1% SDS, and protease inhibitor cocktail (Roche cOmplete™), which was supplemented with 1 mM NaF, 1 mM phenylmethylsulfonyl fluoride, and 1 mM Na3VO4. Whole cell lysate proteins were separated by SDS‐PAGE and electroblotted onto nitrocellulose membranes, which were incubated with several primary antibodies, including NFAT, phospho‐CREB (Abcam), phospho‐NF‐κB, NF‐κB, CREB, phospho‐Stat3, Stat3, phospho‐AKT, AKT, phospho‐p38, p38, phospho‐ERK, ERK (Cell Signaling), phospho‐JNK, JNK (Santa Cruz), and β‐actin (Sigma‐Aldrich). The immunocomplexes were then detected with horseradish peroxidase‐conjugated IgG (Jackson ImmunoResearch Laboratories), and the reaction was developed using an ECL detection kit (Amersham) in an ImageQuant LAS 4000 system (GE Healthcare Life Sciences).
Phospho nf κb
Manufactured by Abcam
Phospho‐NF‐κB is a lab equipment product that detects the phosphorylated form of the NF‐κB protein. NF‐κB is a transcription factor that plays a key role in regulating inflammatory and immune responses in cells. The product enables researchers to measure the activation status of NF‐κB, which is important for understanding cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Imagequant las 4000 system, by GE Healthcare (1 mentions)
Phospho erk, by Cell Signaling Technology (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Ecl detection kit, by Cytiva (1 mentions)
Phospho creb, by Abcam (1 mentions)
Horseradish peroxidase conjugated igg, by Jackson ImmunoResearch (1 mentions)
Stat3, by Cell Signaling Technology (1 mentions)
Phospho stat3, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Goat anti rabbit igg h l hrp, by Cell Signaling Technology (1 mentions)
2 protocols using phospho nf κb
1
Western Blot Analysis of Cell Signaling
2
Hippocampal Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The hippocampal tissues were lysed in RIPA buffer (Cat. No. R0010, Solarbio) containing the phosphatase and protease inhibitors. Protein concentrations were determined by the Bradford method. 10% or 12% SDS-PAGE gel were used to separate the proteins, and then which were transferred to the PVDF membrane (Cat. No. ISEQ00010, Solarbio). The membranes then incubated with primary antibodies diluted in TBST at 4°C overnight, then which were incubated with the HRP-conjugated secondary antibody at room temperature for 1.5–2 h (Liang et al., 2017 (link)). The primary antibodies used were: BDNF (1:1000, Cat. No. ab108319, Abcam), ERK1/2 or phospho-ERK1/2, CREB or phospho-CREB, NFκB or phospho-NFκB (1:2000, Cat. No. 4695, 9101, 4034, 4095, 3033, 3034, Cell Signaling), and the secondary antibodies used were: Goat anti Rabbit IgG (H+L)/HRP (1:5000, Cat. No. ZB2301, Zhongshan JinQiao biotechnology), Goat anti mouse IgG (H+L)/HRP (1:5000, Cat. No. ZB2305, Zhongshan JinQiao biotechnology).
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!