Total protein of mouse lung tissue and cells was extracted using RIP protein lysate containing 1% PMSF. The nuclear and cytoplasmic proteins of HFL1 were extracted using a Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime Biotechnology, Beijing). The protein concentration of each sample was detected by BCA method. The proteins were separated by SDS-PAGE gel electrophoresis, transferred onto PVDF membrane, and sealed with 5% skimmed milk powder at room temperature for 1 h. The PVDF membranes were then incubated overnight with COL1A1 (1:1000, Cell Signaling Technology), α-SMA (1:1000; Cell Signaling Technology), FN (1:1000; Cell Signaling Technology), FOXO3 (1:1000; Abcam), SPON1 (1:1000; Signalway Antibody), Smad3 (1:1000; Affinity), Smad7 (1:1000; Affinity), GAPDH (1:1000; Proteintech), and Lamin B1 (1:1000; Affinity) primary antibody at 4 °C. After washing with TBST three times, secondary antibody (1:10000) was added and the mixture was incubated at room temperature for 2 h. Finally, a chemiluminescence imaging system (Tanon, Shanghai) was used to obtain protein images, and Image J software was used for protein quantification.
Lamin b1
Manufactured by Affinity Biosciences
Sourced in China
Lamin B1 is a structural protein found in the nuclear lamina, a network of proteins lining the inner nuclear membrane. It plays a key role in the organization and stability of the nucleus.
Automatically generated - may contain errors
Lab products found in correlation
Smad3, by Affinity Biosciences (1 mentions)
Smad7, by Affinity Biosciences (1 mentions)
Col1a1, by Cell Signaling Technology (1 mentions)
α sma, by Cell Signaling Technology (1 mentions)
Foxo3, by Abcam (1 mentions)
Gapdh, by Proteintech (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Beyotime (1 mentions)
Lipopolysaccharide lps, by Solarbio (1 mentions)
P erk, by Affinity Biosciences (1 mentions)
P jnk, by Affinity Biosciences (1 mentions)
Bca protein analysis kit, by Meilun (1 mentions)
C myc, by ZenBio (1 mentions)
Cyclin d1, by ZenBio (1 mentions)
Radioimmunoprecipitation assay (ripa), by Meilun (1 mentions)
β catenin, by ZenBio (1 mentions)
Vimentin, by ZenBio (1 mentions)
P iκb, by Affinity Biosciences (1 mentions)
Gapdh, by Affinity Biosciences (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Meilun (1 mentions)
Glyceraldehyde 3 phosphate dehydrogenase gapdh, by Affinity Biosciences (1 mentions)
5 protocols using lamin b1
1
Lung Tissue Protein Extraction and Analysis
2
Protein Expression Analysis in Wound Healing
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After incubation with PBS, OA-RD17 (1 nM), lipopolysaccharide (LPS, 1 μg/mL) (Solarbio, China), specific TLR4 inhibitor (1 μg/mL), miR-632 mimic (50 nM), or miR-632 inhibitor (100 nM) for 24 h, respectively, cell lysates (RIPA: PMSF: phosphatase inhibitor = 100:1:1; RIPA and PMSF, Meilun Biotechnology, Dalian, China; phosphatase inhibitor, Roche, Shanghai, China) were used to extract total protein in keratinocytes and macrophages. Moreover, cell lysates were also used to extract total protein in wound tissues from SD rats. The extracted proteins were quantified using the Bradford method (BCA protein analysis kit, Meilun, Dalian, China). The protein samples were then separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), electro-imprinted on polyvinylidene fluoride membranes, and recorded and analyzed quantitatively using the Bole exposure software system. Primary antibodies, including GAPDH, Lamin B1, P38, P-P38, ERK, P-ERK, JNK, P-JNK, IκB, P-IκB, P65, P-P65 (Affinity, China), GSK3β, β-catenin, Cyclin D1, c-MYC, and Vimentin (ZEN BIO, China) were used following the provided instructions.
3
Carbon Tetrachloride-Induced Liver Injury
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Sch B was purchased from Selleck Chemicals (Houston, TX, USA). Carbon tetrachloride (CCl4) was purchased from Chinasun Specialty Products Co., Ltd. (Changshu, Jiangsu, China). Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit anti-PPARγ, anti-F4/80, anti-alpha smooth muscle actin (α-SMA), anti-CD86, Lamin B1, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were obtained from Affinity Biosciences (Changzhou, Jiangsu, China). Rabbit antibodies against nuclear factor (NF)-κB and phospho-IκBα were purchased from Cell Signaling Technology (Danvers, MA, USA). Secondary antibodies used in western blotting and immunohistochemical staining were purchased from Affinity Biosciences. Secondary antibodies used in immunofluorescence staining were purchased from Abcam (Cambridge, MA, USA).
4
Analyzing Hepatic Lipid Metabolism Markers
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The commercial kits for alanine aminotransferase (ALT), aspartate amino-transferase (AST), malondialdehyde (MDA), oxidized/reduced glutathione (GSSG/GSH), catalase (CAT) and glutathione (GSH) detection were obtained from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Serum total triglycerides (TG) and total cholesterol (TC) kits were purchased from BIOSINO Biotechnology and Science, Inc. (Beijing, China). The TRIzol agent was purchased from Invitrogen, Inc. (Carlsbad, CA, USA). RNA reverse transcription reagents and the SYBR Premix Ex Taq II kits were obtained from TAKARA Bio, Inc. (Otsu, Shiga, Japan). Rabbit monoclonal F4/80 antibody and rabbit monoclonal anti-LC3B were purchased from Cell Signaling Technology (Billerica, MA, USA). Rabbit monoclonal β-actin antibody, rabbit polyclonal Nrf2, Lamin B1, peroxisome proliferator-activated receptor α (PPARα), peroxisome proliferator-activated receptor γ (PPARγ), SREBP-1c, FAS, acyl CoA oxidase 1 (ACOX1), stearoyl-CoA desaturase 1 (SCD1) antibodies, and the secondary antibody were obtained from Affinity Biosciences, Inc. (Cincinnati, OH, USA). All chemicals used were of the highest grade commercially available.
5
Total Protein Extraction and Western Blot
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Total protein of mouse lung tissue and cells was extracted using RIP protein lysate containing 1% PMSF. The nuclear and cytoplasmic proteins of HFL1 were extracted Proteintech), and Lamin B1 (1:1000; Affinity) primary antibody at 4 °C. After washing with TBST three times, secondary antibody (1:10000) was added and the mixture was incubated at room temperature for 2 h. Finally, a chemiluminescence imaging system (Tanon, Shanghai) was used to obtain protein images, and Image J software was used for protein quantification.
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