The collected HUVECs (1 × 105 cells/well) in each group were inoculated into sterile cover glasses in a 6-well plate, respectively. After incubation overnight, HUVECs were fixed by 4% paraformaldehyde (Sigma, St. Louis, MO, USA) and permeated by 0.5%Triton X-100. After sealing with normal goat serum, the HUVECs were exposed with anti-p-TAK1 (1 : 50, Abcam, St. Louis, MO, USA) overnight at 4°C and fluorescent secondary antibody (1 : 100; Abcam, St. Louis, MO, USA) at 37°C for 1 h. After DAPI (Thermo Fisher Scientific, Waltham, MA, USA) nucleation, the fluorescence intensity was observed using a fluorescence microscope (Olympus, CKX41) [29 (link)].
Anti p tak1
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-p-TAK1 is a primary antibody that recognizes the phosphorylated form of the protein TAK1 (transforming growth factor-beta-activated kinase 1). TAK1 is a serine/threonine kinase that plays a critical role in various signaling pathways, including those involved in the regulation of inflammation and immune responses.
Automatically generated - may contain errors
Lab products found in correlation
Ros assay kit, by Beyotime (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modi ed eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Radioimmunoprecipitation assay (ripa), by Beyotime (2 mentions)
Phenylmethanesulfonyl uoride pmsf, by Thermo Fisher Scientific (2 mentions)
Anti p38, by Abcam (2 mentions)
Anti p p38, by Abcam (2 mentions)
Anti erk1 2, by Abcam (2 mentions)
Anti jnk, by Abcam (2 mentions)
Anti p jnk, by Abcam (2 mentions)
Sybr green master mix, by Vazyme (2 mentions)
Hiscript reverse transcriptase, by Vazyme (2 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
Phosphatase inhibitor cocktail, by Beyotime (2 mentions)
Anti p erk1 2, by Abcam (2 mentions)
Anti tak1, by Abcam (2 mentions)
Sb202190, by MedChemExpress (2 mentions)
Anti p nf κb, by Abcam (2 mentions)
Anti nf κb, by Abcam (2 mentions)
4 protocols using anti p tak1
1
Immunofluorescence Imaging of Phosphorylated TAK1
2
Protein Expression Analysis in HUVECs
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Total proteins were extracted with RIPA lysate from the collected HUVECs in each group, and protein concentration was tested by BCA method (Beyotime, China). Protein samples (40 μg) were separated by 10% SDS-PAGE and transferred to PVDF membranes (Roche). PVDF membranes were blocked with 5% skimmed milk powder at room temperature for 2 h and incubated with primary antibody (dilution ratio: 1∶1000) overnight at 4°C and secondary antibody (dilution ratio: 1∶2000) at 37°C for 2 h. The membranes were addressed with ECL solution (Thermo Fisher Scientific), and the results were acquired with a gel imaging system. p-IRAK1 is referenced by IRAK1, p-TAK1 is referenced by TAK1, and GAPDH was applied as the reference for the other proteins. Anti-interleukin-1 receptor-associated kinase 1 (IRAK1), anti-p-IRAK1, anti-TGF-beta-activated kinase 1 (TAK1), anti-p-TAK1, anti-Bax, anti-caspase 3, anti-mitofusion1 (MFN1), anti-mitofusion2 (MFN2) and anti-opticatrophy1 (OPA1) were all purchased from Abcam (USA) [28 (link)].
3
Inflammatory Signaling Pathways in RAW264.7 Cells
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Materials BAC (purity ≥ 98%) and RAW264.7 cells were provided by Yousi Scienti c Co., Ltd., and the Chinese Academy of Sciences (Shanghai, China). LPS from Escherichia coli and TPCK (NF-κB inhibitor) were obtained from Sigma (St. Louis, MO, USA). SB202190 (p38 MAPK inhibitor), U0126 (ERK inhibitor), SP600125 (JNK inhibitor), and Cell-Counting Kit-8 (CCK-8) were obtained from Medchem Express (St.
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
4
Inflammatory Signaling Pathways in RAW264.7 Cells
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Materials BAC (purity ≥ 98%) and RAW264.7 cells were provided by Yousi Scienti c Co., Ltd., and the Chinese Academy of Sciences (Shanghai, China). LPS from Escherichia coli and TPCK (NF-κB inhibitor) were obtained from Sigma (St. Louis, MO, USA). SB202190 (p38 MAPK inhibitor), U0126 (ERK inhibitor), SP600125 (JNK inhibitor), and Cell-Counting Kit-8 (CCK-8) were obtained from Medchem Express (St.
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
Louis, MO, USA). Trizol and phenylmethanesulfonyl uoride (PMSF) were acquired from Ambion (CA, USA) and Aladdin (Shanghai, China). HiScript Reverse Transcriptase and SYBR Green Master Mix were provided by VAZYME (Nanjing, China). ROS Assay Kit, BCA Protein Assay Kit, radioimmunoprecipitation assay (RIPA), and phosphatase inhibitor cocktail were acquired from Beyotime (Shanghai, China).
Dulbecco's modi ed Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY). Anti-NF-κB, anti-JNK, anti-p38, anti-ERK1/2, anti-TAK1, anti-p-NF-κB, anti-p-JNK, anti-p-p38, anti-p-ERK1/2, and anti-p-TAK1 antibodies were purchased from Abcam (Cambridge, UK). Anti-IκBα and anti-p-IκBα antibodies were obtained from A nity Bio Reagents (Golden, CO, USA).
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