We carried out the western blot analysis based on the standard procedure in previous reports. The dorsal root ganglion tissues were homogenized in cold RIPA lysis buffer with protease inhibitors. After centrifugation at 10,000 g for 20 min, the supernatant was collected for protein quantification. 10 μg of protein was separated by 10% SDS-PAGE and transferred onto the PVDF membrane (Roche). Then, the blot was blocked with defatting milk powder (5%) at room temperature for one hour. After that, the blot was incubated overnight with the following primary antibodies: anti-IL-6 (1 : 1000, Proteintech, USA), anti-MAPK1 (1 : 200, Proteintech, USA), anti-TP53 (1 : 200, Proteintech, USA), and anti-GAPDH (1 : 1000, Proteintech, USA). After incubation, the blots were incubated with horseradish peroxidase-labeled secondary antibody for 60 min at room temperature. Following incubation, the blots were measured by a chemiluminescence reagent (PerkinElmer, USA), and the band intensity quantification was performed using ImageJ software (NIH). GAPDH was used as an endogenous control.
Anti tp53
Manufactured by Proteintech
Sourced in United States
The Anti-TP53 product is an antibody that specifically binds to the TP53 protein. TP53 is a transcription factor that plays a crucial role in regulating cell growth and division. The Anti-TP53 antibody can be used in various laboratory techniques to detect and analyze the TP53 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Proteintech (3 mentions)
Chemiluminescence reagent, by PerkinElmer (1 mentions)
Pvdf membrane, by Roche (1 mentions)
Anti il 6, by Proteintech (1 mentions)
Ab124964, by Abcam (1 mentions)
Ab45688, by Abcam (1 mentions)
Ab124805, by Abcam (1 mentions)
Ab109520, by Abcam (1 mentions)
Anti p p38, by Cell Signaling Technology (1 mentions)
Anti mdm2, by Cell Signaling Technology (1 mentions)
Ecl kit, by 4A Biotech (1 mentions)
Anti col1, by Cell Signaling Technology (1 mentions)
Anti cd68, by Proteintech (1 mentions)
Anti t p38, by Cell Signaling Technology (1 mentions)
Anti vimentin, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Proteintech (1 mentions)
Anti cdkn2a, by Proteintech (1 mentions)
Anti dnmt3a, by Proteintech (1 mentions)
Anti dnmt3b, by Proteintech (1 mentions)
Anti dnmt1, by Proteintech (1 mentions)
4 protocols using anti tp53
1
Western Blot Analysis of Neuroinflammation
2
Western Blot Analysis of Fibrosis Markers
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We performed western blot (WB) analysis as previously described36 using an ECL kit (4AW011; purchased from 4A Biotech Co., Ltd) and the following antibodies: anti‐GAPDH (1:5000, 60004‐1‐Ig; Proteintech, USA), anti‐αSMA (1:1000, ab124964; Abcam, UK), anti‐fibronectin (1:1000, ab45688; Abcam, UK), anti‐MDM2 (1:1000, #86934; Cell Signaling Technology, USA), anti‐t‐p53 (1:1000, 10442‐1‐AP; Proteintech, USA), anti‐Col1 (1:1000, #84336; Cell Signaling Technology, USA), anti‐vimentin (1:1000, #5741; Cell Signaling Technology, USA), anti‐CD68 (1:5000, 25747‐1‐AP; Proteintech, USA), anti‐p‐p38 (1:1000, #4511; Cell Signaling Technology, USA), anti‐t‐p38 (1:1000, #8690; Cell Signaling Technology, USA), anti‐p21 (1:1000, ab109520; Abcam, UK), and nti‐FSP1 (1:1000, ab124805; Abcam, UK).
3
Protein Expression Analysis in Cancer Cells
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Total protein was extracted from SCL-1 and A431 cells using Total Protein Extraction Kit (Solarbio). Protein samples were separated by polyacrylamide gel electrophoresis, and then electro-blotted onto polyvinylidene fluoride membranes (Merck Millipore, Billerica, MA, USA). The membranes were incubated with primary antibody anti-CDKN2A (1:1000, Proteintech, Wuhan, China), anti-TP53 (1:2000, Proteintech), anti-DNMT1 (1:1000, Proteintech), anti-DNMT3A (1:1000, Proteintech), anti-DNMT3B (1:1000, Proteintech), or anti-β-actin (1:5000, Proteintech) overnight at 4°C. After washed with Tris Buffered saline Tween, the membranes were incubated with horseradish peroxidase-conjugated secondary antibody (1:5000, Proteintech). WB signals were analyzed by Image J software.
4
Investigating Cytoskeletal Protein Regulation
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After cells were seeded overnight, TPM1, TPM2, TPM3, YAP1, and TP53 siRNA (Tsingke) and Lipofectamine RNAiMAX (Invitrogen) were mixed for transfection on the following day. Control cells were transfected with scrambled siRNA (Tsingke). The knockdown efficiency was verified on the 3rd day, and the cells were then used for further experiments. For immunoblotting, cells were lysed in radioimmunoprecipitation assay buffer supplemented with 1% phenylmethanesulfonyl fluoride, and protein quantification was performed using the bicinchoninic acid method. The extracted proteins were separated by 10% gel electrophoresis and transferred to polyvinylidene fluoride membranes. The membranes were incubated with the following primary antibodies at 4 °C overnight: anti-GAPDH (Proteintech, dilution 1:50 000), anti-BAX (Proteintech, dilution 1:2 000), anti-Caspase-3 (Proteintech, dilution 1:1 000), anti-Caspase-9 (CST, dilution 1:1 000), anti-TPM1 (Proteintech, dilution 1:1 000), anti-TPM2 (Proteintech, dilution 1:1 000), anti-TPM3 (Proteintech, dilution 1:500), anti-YAP1 (Santa Cruz, dilution 1:500), anti-p-YAP1 (CST, dilution 1:1 000), and anti-TP53 (Proteintech, dilution 1:1 000). Primary antibody binding was detected using enhanced chemiluminescence with appropriate horseradish peroxidase-conjugated secondary antibodies.
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