After treatment, cells were collected and lysed in RIPA buffer. The protein concentrations were measured by Bradford protein assay kit (Sigma-Aldrich). Equal amount of protein was loaded and subjected to SDS-PAGE and transferred to PVDF membrane (Millipore, Boston, MA, USA). After blocking with 5% skimmed milk for 1 h at room temperature, PVDF membrane was incubated with primary antibody overnight at 4°C. The following antibodies were used in our study: anti-caspase-3, anti-caspase-9, anti-Bcl-2, anti-Bcl-xl, anti-Mcl-1, anti-Bid, anti-Bax, anti-cytochrome c, anti-Smac/DIABLO, anti-Bax (6A7), anti-ATF4, and anti-CHOP were purchased from Cellular Signaling Technology (Danvers, MA, USA) and anti-p-PERK, anti-PERK, anti-p-eIF2α, and anti-GAPDH (Abcam, San Diego, CA, USA) Then, the membrane was incubated with secondary antibody and visualized by ECL (Thermo Scientific, Rockford, USA). Purification of cytosolic fraction and Bax immunoprecipitation were performed as described earlier [9 (link)].
Anti p perk
Manufactured by Abcam
Sourced in United States
Anti-p-PERK is a lab equipment product that detects and quantifies the phosphorylated form of the Protein Kinase R (PKR)-like Endoplasmic Reticulum Kinase (PERK) protein. It is used for research purposes to study the activation of PERK, a key regulator of the unfolded protein response.
Automatically generated - may contain errors
Lab products found in correlation
Anti p erk, by Abcam (2 mentions)
Anti gapdh, by Abcam (2 mentions)
Anti p eif2α, by Abcam (1 mentions)
Bradford protein assay kit, by Merck Group (1 mentions)
Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Enhanced chemiluminescent detection system, by Bio-Rad (1 mentions)
Anti p ire1α, by Abcam (1 mentions)
Protein extraction reagent, by Beyotime (1 mentions)
Anti jnk, by Abcam (1 mentions)
Anti p jnk, by Abcam (1 mentions)
Hrp labeled goat anti rabbit igg secondary antibody, by Beyotime (1 mentions)
Bca protein quantification kit, by Abcam (1 mentions)
Anti cleaved caspase 3, by Abcam (1 mentions)
Anti grp78, by Abcam (1 mentions)
2 protocols using anti p perk
1
Western Blot Analysis of Apoptotic Markers
2
ER Stress Pathway Activation in Lung Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Right upper lung tissues and A549 cells were lysed with protein extraction reagent (P0013; Beyotime Institute of Biotechnology) according to the manufacturer's protocol. Protein concentration was determined using BCA protein quantification kits (ab102536; Abcam). Proteins (30 µg proteins in each group) were separated by 12% gradient SDS-PAGE and were transferred to polyvinylidene fluoride membranes. Nonspecific binding was blocked with 10% non-fat dry milk in Tris-buffered saline (TBS) at room temperature for 1 h. The membranes were then incubated overnight at 4°C with anti-PERK (1:1,000 dilution), anti-p-PERK (1:1,000 dilution), anti-GRP78 (1:1,000 dilution), anti-JNK (1:1,000 dilution), anti-p-JNK (1:1,000 dilution), anti-CHOP (1:1,000 dilution), anti-IRE1α (1:1,000 dilution), anti-p-IRE1α (1:1,000 dilution), anti-p-50ATF6α (1:1,000 dilution), anti-cleaved caspase-3 (1:1,000 dilution) and anti-GAPDH (1:1,000 dilution; ab8245; Abcam). After washing with TBS containing 20% Tween-20 (20% Tween-20: TBS =2.6:1,000), the membranes were incubated with an HRP-labeled goat anti-rabbit IgG secondary antibody (1:7,500 dilution; A0208; Beyotime Institute of Biotechnology) at room temperature for 1.5 h. The blots were visualized using an enhanced chemiluminescent detection system (Bio-Rad, Hercules, CA, USA).
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