Protein expression in BMSCs was determined by immunoblotting as previously described [27 (link)]. Anti-LRRc17 (1:1000, Proteintech), anti-p21 (1:1000, Affinity), anti-p16 (1:1000, CST), anti-p53 (1:1000, CST), anti-mTOR (1:500, Affinity), anti-phospho-mTOR (1:500, CST), anti-PI3K (1:1000, CST), anti-phospho-PI3K (1:1000, Affinity), anti-P62 (1:5000, ABclonal), anti-LC3B (1:1000, CST), anti-Beclin1 (1:1000, CST), anti-OPA1 (1:1000, CST), anti-Drp1 (1:1000, CST), anti-JNK (1:1000, Abclonal) abti-phosphor-JNK (1:1000, Abclonal) and anti-Calpain1 (1:1000, Abclonal) antibodies were used to analyze protein expression. The signal was visualized using an Immobilon Western Chemiluminescent HRP Substrate Kit (Millipore, USA). Quantification of the gray values of the bands was performed with ImageJ software.
Anti p21
Manufactured by Affinity Biosciences
Sourced in United States, China
Anti-p21 is a research antibody that binds to the p21 protein, which is a cyclin-dependent kinase inhibitor involved in cell cycle regulation. It can be used to detect and quantify the expression of p21 in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti p53, by Affinity Biosciences (3 mentions)
Immobilon western chemiluminescent hrp substrate kit, by Merck Group (1 mentions)
Anti p62, by ABclonal (1 mentions)
Anti mtor, by Affinity Biosciences (1 mentions)
Anti phospho pi3k, by Affinity Biosciences (1 mentions)
Anti jnk, by ABclonal (1 mentions)
Beyoecl plus developer, by Beyotime (1 mentions)
Ripa cleavage buffer, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Anti β actin, by Affinity Biosciences (1 mentions)
Anti β tubulin, by Affinity Biosciences (1 mentions)
Anti tnf α, by Affinity Biosciences (1 mentions)
Mouse anti cd31, by Wuhan Servicebio Technology (1 mentions)
Anti nf κb, by Affinity Biosciences (1 mentions)
Anti il 6, by Wuhan Servicebio Technology (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Anti nrf2, by ABclonal (1 mentions)
Anti ho 1, by ABclonal (1 mentions)
4 protocols using anti p21
1
Protein Expression Analysis in BMSCs
2
Protein Expression Analysis in D283 and D341 Cells
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D283 and D341 cells were lysed with protease inhibitor, and RIPA cleavage buffer (Thermo Fisher Scientific, Shanghai, China). The protein lysates were resolved via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and imprinted on polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, USA) for analysis. Anti-CENEP (1:1,000 dilution, DF7745; Affinity Biosciences, OH, USA), anti-P53 (1:500 dilution, AF0879; Affinity Biosciences, OH, USA), anti-P21 (1:500 dilution, AF6290; Affinity Biosciences, OH, USA), anti-CDK1 (1:500 dilution, DF6024; Affinity Biosciences, OH, USA) and anti-β-Actin (1:5,000 dilution, AF7018; Affinity Biosciences, OH, USA) were incubated overnight. Goat Anti-Rabbit IgG (H+L) HRP-conjugated secondary antibodies (1:5,000 dilution; S0001, Affinity Biosciences, OH, USA) were added at room temperature for 2 h. The membrane was detected with BeyoECL Plus developer (Beyotime, Shanghai, China).
3
Assessing 786-O Cell Behavior
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The Colony Formation, Transwell Migration, and Invasion assay of 786-O cell were performed as described previously (13 (link)). A minimum of five random fields of view were immediately captured on the 786-O cell. For western blot assay, 786-O cell were incubated with anti-DUSP1 (Cat# AF5286, Affinity Biosciences), anti-β-tubulin (Cat# T0023, Affinity Biosciences), anti-p21 (Cat# AF6290, Affinity Biosciences) and anti-p53 (Cat# AF0879, Affinity Biosciences), following are the specific methods shown (14 (link)).
4
Immunohistochemical Analysis of Aortic Proteins
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Paraffin slices of the aortas were deparaffinized, rehydrated, and incubated in goat serum for 1 h at 37 °C. Rabbit anti-p16 (1:300, Affinity Biosciences, Changzhou, China), anti-P21 (1:300, Affinity Biosciences, Changzhou, China), anti-P53 (1:300, Affinity Biosciences, Changzhou, China), anti-SIRT1 (1:100, Boster, Wuhan, China), anti-Nrf2 (1:150, Abclonal, Wuhan, China), anti-HO-1 (1:100, Abclonal, Wuhan, China), anti-IL-6 (1:300, Servicebio, Wuhan, China), anti-TNF-α (1:200, Affinity Biosciences, Changzhou, China), anti-NF-κB (1:200, Affinity Biosciences, Changzhou, China), and mouse anti-CD31 (1:300, Servicebio, Wuhan, China) antibodies were first added and incubated overnight to determine the contents of these proteins in the aortas at 4 °C. After that, FITC-labelled anti-rabbit and anti-mouse IgG antibodies were used as secondary antibodies (Servicebio, Wuhan, China) for detection. One hour later, DAPI was used for nuclear counterstaining. Finally, images were obtained with a fluorescence microscope (Leica, Wetzlar, Germany). ImageJ software was used to quantify the results.
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